Ncv ns3 protease inhibitors
Abstract
A compound of formula (I): wherein: the ipiq 1; R 1 is CONR10SO2R6; R2 is C1-C6 alkyl or C2-C6 alkenyl, wherein said alkyl or alkenyl optionally substituted with 1 to 3 halo; R 3 is C 1 -C 8 alkyl or C3-C8 cycloalkyl; R5 is H; R6 is C3-C6 cycloalkyl, Y is C (= 0); Z is O; M is C1-C12 alkylene or C2-C12 alkenylene; each entry R10 is independently H or C1-C6 alkyl; or a pharmaceutically acceptable salt or hidrat.Prijava contains 16 claims.

Term
No projected expiry on record.
- Priority
- Filed
- Granted
- Today
17 claims: 4 independent, 13 dependent
- 1Compound of formula (I):1. Jedinjenje formule (I): characterized in that: naznačeno time što: i p i q su 1;R1 je CONR10SO2R6;ipiq su 1;R1 is CONR10SO2R6;R2 is C 1 -C 6 alkyl or S2-Sb alkenyl, said alkyl or alkenyl being optionally substituted with 1 to 3 halo;R2 je СгСб alkil ili С2-Сб alkenil, pri čemu je navedeni alkil ili alkenil izborno supstituisan sa 1 do 3 halo;R3 is C1-Cs alkyl or C3-Cs cycloalkyl;R3 je C|-Cs alkil ili C3-Cs cikloalkil;RNo 5jeH;RЧ 5jeH;R6jeC3-C6 cikloalkil;R6jeC3-C6 cycloalkyl;Y is C (= O);YjeC(=O);Z is 0;Zje 0;M is C1-C12 alkylene or C2-Ci2 alkenylene;and each R10 is independently H or C 1 -C 6 alkyl;or a pharmaceutically acceptable salt or hydrate thereof. M je C|-C]2 alkilen ili C2-Ci2 alkenilen;i svako R10 je nezavisno H ili Cj-Сб alkil;ili njegova farmaceutski prihvatljiva so ili hidrat.
- 10A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1-9 or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier. 10. Farmaceutska kompozicija naznačena time što sadrži efikasnu količinu jedinjenja prema bilo kom od patentnih zahteva 1-9 ili njegove farmaceutski prihvatljive soli ili hidrata, i farmaceutski prihvatljiv nosač. 51101Β 51101Β
- 14Use of a compound according to any one of claims 1-9, or a pharmaceutically acceptable salt or hydrate thereof, in the preparation of a medicament for the prevention or treatment of HCV infection in a subject in need thereof. 14. Primena jedinjenja prema bilo kom od patentnih zahteva 1-9 ili njegove fannaceutski prihvatljive soli ili hidrata, u pripremi leka za prevenciju ili lečenje HCV mfekcije kod subjekta kod koga postoji potreba za tim.
Independent claims4
503 paragraphs in 25 sections, as filed
The present invention relates to macrocyclic compounds useful as inhibitors of hepatitis C virus (HCV) NS3 protease, to their synthesis, and to their use in the treatment or prevention of HCV infection.
BACKGROUND OF THE INVENTION Hepatitis C virus (HCV) infection is a major health problem leading to chronic liver disease, such as cirrhosis and hepatocellular carcinoma. in a significant number of infected persons, which is estimated to represent 2-15% of the world's population. It is estimated that there are 3.9 million infected people in the United States alone, according to the US Centers for Disease Control, which is approximately five times the number of people infected with the human immunodeficiency virus (HIV). According to the World Health Organization, more than 170 million people are infected worldwide, with at least 3 to 4 million people infected each year. After infection, about 20% of people lose the virus. but the rest struggled with HCV for the rest of their lives. Ten to twenty percent of chronically infected people eventually develop cirrhosis or cancer that destroys the liver. Viral disease is transmitted parenterally through infected blood and blood products, infected needles, sexually or vertically from infected mothers or mothers who are carriers of the virus to their offspring.
Current treatments for HCV infection, which are limited to immunotherapy with recombinant interferon-α alone or in combination with the nucleoside analogue ribavirin, have limited clinical benefits. In addition, no HCV vaccine has been established. Consequently, there is an urgent need for improved therapeutic agents that effectively fight chronic HCV infection. The current state of the art in the treatment of HCV infection is discussed in the following literature: B. Dymock, et al., Novel approaches to the treatment of hepatitis C virus infection, Antiviral Chemistry & Chemotherapy, 11: 79-96 (2000): H. Rosen, et al., Hepatitis C virus: current understanding and prospects for future therapies .
51101 Β
Molecular Medicine Today, 5: 393-399 (1999); D. Moradpour, et a! .. Current and evolving therapies for hepatitis C, European J. Gastroenterol. Hepatol., 11: 1189-1202 (1999); R. Bartenschlager, Candidate Targets for Hepatitis C Virus-Specific Antiviral Tcegar. Intervirology, 40: 378-393 (1997); GM Lauer and BD Walker, Hepatitis C Virus lnfection, N. Engl. J. Med., 345: 41-52 (2001): BW Dvmock, Emerging therapies for hepatitis C virus infection, Emerging Drugs. 6: 13-42 (2001); go. Crabb, Hard-Won Advances Spark Excitement about Hepatitis C, Science: 506-507 (2001).
Several enzymes encoded by the virus have been hypothesized as target sites for therapeutic intervention, including metalloprotease (NS2-3). serine protease (NS3). helicase (NS3), and RNA-dependent RNA polymerase (NSSB). NS3 protease is located in the Nterminal domain of the NS3 protein and is considered the first target site for the drug as it is responsible for intramolecular separation at the NS3 / 4A site and for downstream intermolecular processing at compounds NS4A / 4B, NS4B / 5A and NS5A / 5B . Previous research has identified classes of peptides, such as hexapeptides as well as tripeptides discussed in U.S. Patent Application US2005 / 0020503. US2004 / 0229818 and US2004 / 00229776, showing degrees of activity in inhibiting NS3 protease. International Application WO 03/064455 discloses macrocyclic peptides that are useful as HCV NS3 protease inhibitors. The object of the present invention is to provide additional compounds which show activity against UCV NS3 protease.
SUMMARY OF THE INVENTION
The present invention relates to novel macrocyclic compounds of formula (I) and / or their pharmaceutically acceptable salts or hydrates. These compounds are useful in inhibiting HCV (hepatitis C virus) NS3 (non-structural 3) proteases, preventing or treating one or more 11 symptoms of HCV infection, either as compounds, their pharmaceutically acceptable salts or hydrates (where appropriate). or as ingredients of a pharmaceutical composition. either in combination or not with other HCV antivirals, anti-infectives, immunomodulators, antibiotics or vaccines. Especially. the present invention relates to a compound of formula (I) and / or a pharmaceutically acceptable salt or hydrate thereof:
51101 Β
<img file="RS51101B_D0001.tif" />
where:
and ρ iq are 1;
R'jeCONR<sup>l0</sup>SO<sub>2</sub>R<sup>6</sup>;
R<sup>2</sup> is C1-C<sub>6</sub> alkyl or C<sub>2</sub>-C<sub>6</sub> alkenyl; wherein said alkyl or alkenyl is optionally substituted with 1 to 3 halo;
R<sup>.</sup> is C1-C<sub>8</sub> alkyl or C<sub>3</sub>-C<sub>8</sub> cycloalkyl;
R<sup>5</sup> is II;
R<sup>6</sup> is S<sub>3</sub>-Sb cycloalkyl;
Y is C (O);
ZjeO;
M is C1-C12 alkylene or C1-C1<sub>2</sub> alkenylene; and each R<sup>10</sup> is independently H or C1-C<sub>6</sub> alkyl.
The present invention also encompasses pharmaceutical compositions comprising a compound of the present invention and methods for preparing such pharmaceutical compositions. The present invention further relates to methods for treating or preventing one or more symptoms of HCV infection.
Other variants. aspects and features of the present invention are either further described in, or will be apparent from, the following description. examples and attached patent claims.
51101Β
DETAILED DESCRIPTION OF THE INVENTION
The present invention encompasses compounds of formula I from the preceding test, and pharmaceutically acceptable salts and / or hydrates thereof. These compounds and their pharmaceutically acceptable salts and / or hydrates are HCV protease inhibitors (e.g., HCV NS3 protease inhibitors). The present invention also encompasses compounds of formulas P and III wherein all are variables as defined for formula L
<img file="RS51101B_D0002.tif" />
<img file="RS51101B_D0003.tif" />
A first embodiment of the present invention is a compound of formula I, II or III, or a pharmaceutically acceptable salt or hydrate thereof, wherein R<sup>1</sup> equal to CONHSCHR<sup>6</sup>, and all other variables are as originally defined (i.e., as defined in the Summary of the Invention). In a first aspect of the first variant, R<sup>1</sup> is CONHSChR<sup>6</sup> where R<sup>6</sup> equals C3-C5 cycloalkyl; and all other variables are as defined in the first embodiment. In the characteristic of the first aspect of the first variant, R<sup>1</sup> is CONIISCPR<sup>6</sup> gdc is R<sup>6</sup> equals cyclopropyl; and all other variables are as defined in the first variant.
Another variant of the present invention is a compound of formula I, II or III, or a pharmaceutically acceptable salt or hydrate thereof, wherein R 1 is C 1 -C<sub>6</sub> alkyl or C<sub>2</sub>-C<sub>6</sub> alkenyl; and all other variables are as originally defined in any of the previous variants. In the first aspect of the second variant, R 1 is C 1 -C<sub>4</sub> alkyl or C<sub>2</sub>-C<sub>4</sub> alkenyl; and all other variables are as originally defined or as defined in any of the preceding embodiments. In another aspect of the second variant, R<sup>2</sup> is C<sub>2</sub>-C<sub>4</sub> alkenyl; and all other variables are as originally defined or as defined in any of the preceding variants. In a characteristic of another aspect of the second variant, R is viml; and all other variables are as defined in the third variant or as defined in any of the preceding variants. In the third aspect of the second variant. R 'is C1-C<sub>4</sub> alkyl; and all others
51101Β variables are as originally defined or as defined in any of the preceding embodiments. In a characteristic of the third aspect of the third extract, R is ethyl; and all other variables are as defined in the third embodiment or as defined in any of the preceding embodiments.
A third embodiment of the present invention is a compound of formula I, II or III, or a pharmaceutically acceptable salt or hydrate thereof. where R<sup>3</sup> equally C3-C8 cycloalkyl or C1-C1 alkyl; and all other variables are as originally defined or as defined in any of the preceding variants. In the first aspect of the third variant, R<sup>3</sup> is C5-C7 cycloalkyl or C1-C8 alkyl; and all other variables are as defined in the third embodiment or as defined in any one of the preceding claims. In another aspect of the third embodiment, R is C5-C<sub>ft</sub> cycloalkyl or C 1 -C 6 alkyl; and all other variables are as defined in the third embodiment or as defined in any one of the preceding claims. In a third aspect of the third embodiment, R is propyl or butyl; and all other variables are as defined in the third embodiment or as defined in any of the preceding embodiments. In a characteristic of the third aspect of the third variant, R<sup>.</sup> is i-propyl, n-butyl or t-butyl; and all other variables are as defined in the third embodiment or as defined in any of the preceding embodiments.
·?
In a fourth aspect of the third embodiment, R is cyclopentyl or cyclohexyl; and all other variables are as defined in the third variant or as defined in any of the preceding variants.
A fourth embodiment of the present invention is a compound of formula I, IIa or IIIa, or a pharmaceutically acceptable salt or hydrate thereof, wherein M is C 1 -C 6 alkylene or C 2 -C 10 alkenylene (including straight or branched chain alkylene or alkenylene); and all other variables are as originally delimited or as defined in any of the preceding variants. In a first aspect of the fourth embodiment, M is C 1 -C 6 alkylene or C<sub>3</sub>-Cx alkenylene (including straight-chain and garnetted chain alkylene or alkenylene); and all other variables are as originally defined or as defined in any of the preceding claims. In another aspect of the fourth embodiment, M is C<sub>4</sub> alkylene or C<sub>4</sub> alkenylene (including straight-chain and garnetted chain alkylene or alkenylene); and all other variables are as defined in the fourth variant or as defined in any of the preceding variants. In a third aspect of the fourth embodiment, M is C 5 alkylene or C<sub>5</sub> alkenylene (including straight and garnetted alkylene or alkenylene); and all other variables are as defined in the fourth variant or as defined in any of the preceding variants. In a fourth aspect of the fourth variant, M is C<sub>6</sub> alkylene or C (> alkenylene) (including straight chain and branched chain alkylene or alkenylene), and all other variables are as defined in
51101 Β the fourth variant or as defined in any of the previous variants. In the fifth aspect of the fourth variant, M is C<sub>7</sub> alkylene or alkenylene (including straight-chain and garnetted chain alkylene or alkenylene); and all other variables are as defined in the fourth variant or as defined in any of the preceding variants. In a sixth aspect of the fourth embodiment, M is (alkylene or C 5 alkenylene) (including straight chain or branched chain alkylene or alkenylene), and all others are variable as defined in the fourth embodiment or as defined in any of the preceding embodiments. In a seventh aspect of the fourth embodiment, M is C 1-4 alkylene or C 1-6 alkenylene (including straight and garnetted alkylene or alkenylene); and all other variables are as defined in the fourth variant or as defined in any of the preceding variants. In an eighth aspect of the fourth embodiment, M is C 10 alkylene or C 10 alkenylene (including straight chain and garnetted alkylene or alkenylene); and all other variables are as defined in the fourth embodiment or as defined in any one of the preceding claims. In the ninth aspect of the fourth variant, M is selected from the following; and all other variables are as defined in the fourth variant or as defined in any of the preceding variants.
A fifth embodiment of the present invention is a compound. or a pharmaceutically acceptable salt or hydrate thereof, selected from the group consisting of the following compounds.
51101 Β
<img file="RS51101B_D0004.tif" />
W-1 W-2 W-21
<img file="RS51101B_D0005.tif" />
W-5 W-8 W-9
51101 Β
<img file="RS51101B_D0006.tif" />
W-40 W-54
51101 Β
<img file="RS51101B_D0007.tif" />
51101Β
<img file="RS51101B_D0008.tif" />
<img file="RS51101B_D0009.tif" />
ΠΙ-96
<img file="RS51101B_D0010.tif" />
<img file="RS51101B_D0011.tif" />
ΤΠ-79
W-78
<img file="RS51101B_D0012.tif" />
ΙΠ-112
<img file="RS51101B_D0013.tif" />
<img file="RS51101B_D0014.tif" />
W-117
W-133
51101Β
<img file="RS51101B_D0015.tif" />
W-136 W-137
<img file="RS51101B_D0016.tif" />
<img file="RS51101B_D0017.tif" />
W-152 W-153
<img file="RS51101B_D0018.tif" />
<img file="RS51101B_D0019.tif" />
<img file="RS51101B_D0020.tif" />
<img file="RS51101B_D0021.tif" />
<img file="RS51101B_D0022.tif" />
<img file="RS51101B_D0023.tif" />
<img file="RS51101B_D0024.tif" />
51101Β
<img file="RS51101B_D0025.tif" />
W-197 W-198 W-199
<img file="RS51101B_D0026.tif" />
W-200 W-201 W-202
<img file="RS51101B_D0027.tif" />
<img file="RS51101B_D0028.tif" />
W-225
W-225
Other embodiments of the present invention pertain to the following;
(a) A pharmaceutical composition comprising an effective amount of a compound of formula I, N or III and a pharmaceutically acceptable carrier.
(b) The pharmaceutical composition according to (a). further comprising another therapeutic agent selected from the group consisting of an HCV antiviral agent, an immunomodulator, and an anti-infective agent.
51101 C (c) The pharmaceutical composition according to (b), wherein the IICV antiviral agent is an antiviral agent selected from the group consisting of an HCV protease inhibitor and an HCV NS5B polymerase inhibitor.
(d) A pharmaceutical combination which is (i) a compound of formula I, II or III and (ii) another therapeutic agent selected from the group consisting of an IIC V antiviral agent, an immunomodulator and an anti-infective agent; wherein both the compound of formula I, I-a or III and the other therapeutic agent are used in an amount that makes the combination effective to inhibit HCV NS3 protease, or to treat or prevent HCV infection.
(e) The combination according to (d), wherein the HCV antiviral agent is an antiviral agent selected from the group consisting of an HCV protease inhibitor and an HCV NS5B polymerase inhibitor.
(f) A method of inhibiting HCV NS3 protease in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula I, II or III.
(g) A method for preventing or treating HCV infection in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula I, II or I.
(h) The method of (g), wherein the compound of formula I, II or III is administered in combination with an effective amount of at least one other therapeutic agent selected from the group consisting of an HCV antiviral agent, an immunomodulator, and an anti-infective agent.
(i) The method of (h), wherein the HCV antiviral agent is selected from the group consisting of an HCV protease inhibitor and an HCV NS5B polymerase inhibitor.
(j) A method of inhibiting HCV NS3 protease in a subject in need thereof, comprising administering to the subject a pharmaceutical composition according to (a), (b) or (c) or a combination according to (d) or (e).
(k) A method of preventing or treating a FICV infection in a subject in need thereof, comprising administering to the subject a pharmaceutical composition according to (a), (b) or (c) or a combination according to (d) or (c).
The present invention also encompasses a compound of the present invention (i) for use in, (ii) for use as a medicament for, or (iii) for use in the preparation of a medicament for: (a) inhibiting HCV NS3 protease, or (b) preventing or treating HCV infections. In these applications, the compounds of the present invention may optionally be used in combination with one or more other therapeutic agents selected from IICV antiviral agents. antiinfectives and immunomodulators.
51101 Β
Additional variants of the invention relate to the pharmaceutical compositions, combinations and methods set forth in (a) - (k) above and the uses set forth in the preceding paragraph, wherein the compound of the present invention is a compound of one of the variants, aspects, classes used herein. , a subclass, or characteristic of the compounds described above. In all these variants. the compound may optionally be used in the form of a pharmaceutically acceptable salt or hydrate as appropriate.
As used herein, the term alkyl means any straight or branched chain alkyl group having the number of carbon atoms in the indicated range. In that way. for example, “C 1-6 alkyl (or C 1 -C 6 alkyl) means all hexyl alkyl and pentyl alkyl isomers as well as n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl. As another example, C 1-4 alkyl means n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl.
The term haloalkyl means an alkyl group in which a hydrogen atom is replaced by a halogen. The term alkoxy means an alkyl-O- group.
The term alkylene means any straight or branched chain alkylene group having the number of carbon atoms in the indicated range. Thus, for example, alkylene- denotes each of C1 to C<sub>6</sub> straight or garnetted alkylenes. A class of alkylene of particular interest in connection with the invention is - (CH<sub>2</sub>)|.<sub>6</sub>-, and sub-classes of particular importance include - (CH<sub>2</sub>) i_<sub>4</sub>- - (CH<sub>2</sub>)].<sub>3</sub>-, - (CH<sub>2</sub>) i-<sub>2</sub>- and -CH<sub>2</sub>-. Also important is aikilene -CH (CH<sub>3</sub>)-.
The term alkenylene means any straight-chain or branched chain of a divalent alkenylene group having a number of carbon allomes in the indicated range.
The term cycloalkyl means any cyclic ring of an alkane or alkene having a number of carbon atoms in the indicated range. In that way. for example. C<sub>3</sub>.s cycloalkyl (or C<sub>3</sub>cycloalkyl ”) means cyclopropyl cyclobutyl. cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. The term cycloalkoxy means a cycloalkyl-O- group.
The term halogen (or halo) means fluorine, chlorine, bromine and iodine (alternatively designated as fluoro, chloro, bromo and iodo).
Unless explicitly stated otherwise, all bands listed herein are included. For example. a heteroaryl ring described to contain from 1 to 3 heteroatoms means that the ring may contain 1, 2 or 3 heteroatoms. It is also to be understood that each range cited herein includes within its scope all sub-ranges within that range. Oxidized forms of the N and S heteroatoms are also included within the scope of the present invention.
When any variable (e.g. R<sup>10</sup>) occurs in more than one pool in any component or in formula I, P or III. or in any other formula depicting and describing the compounds according to the invention, its definition is in any case independent of its
51101 Β definitions in every other case. Also, combinations of substituents and / or variables are permitted only if such combinations result in stable compounds.
Unless explicitly stated otherwise, substitution with said substituent is allowed on any ring atom (e.g., aryl, heteroaromatic ring, or saturated heterocyclic ring) provided that such ring substitution is chemically permissible and results in a stable compound. A stable compound is a compound that can be prepared and isolated and whose structure and properties remain or can be caused to remain substantially unchanged for a period of time sufficient to allow the compound to be used for the purposes described herein (e.g., therapeutic or prophylactic use). of the subject).
As a result of the choice of substituents and substituent patterns, certain compounds of those of the present invention may have asymmetric centers and may occur as mixtures of stereoisomers, or as individual diastereomers or enantiomers. All isomeric forms of these compounds, whether isolated or in mixtures, are within the scope of the present invention.
As may be apparent to one skilled in the art, certain compounds of those of the present invention may exist as tautomers. For the purposes of the present invention, a reference to a compound of formula I, P, or III is a reference to the compound per se, or to any of its tautomers per se, or to mixtures of two or more tautomers.
The compounds of the present invention are useful in inhibiting HCV protease (e.g., HCV NS3 protease) and preventing or treating HCV infection. For example, the compounds of the present invention are useful in the treatment of HCV infection as there is a suspicion of past exposure to HCV virus through for example blood transfusions, body fluid exchange, bites, accidental needle sticks or blood exposures of a patient during surgery.
The compounds of the present invention are useful in preparing and performing screening tests for antiviral compounds. For example. the compounds of the present invention are useful for isolating enzyme mutants, which are an excellent screening agent for stronger antiviral compounds. In addition, the compounds of the present invention are useful in establishing or determining the binding site of other antiviral agents to HCV protease, e.g., by competitive inhibition. Thus, the compounds of the present invention are commercial products to be sold for these purposes.
The compounds of the present invention may be administered in the form of pharmaceutically acceptable salts. The term pharmaceutically acceptable salt means a salt which possesses the efficacy of the parent compound and which is not biologically or otherwise undesirable
51101Β (e.g., not toxic, or otherwise harmful to the recipient). Suitable salts include acid addition salts which may, for example, be formed by mixing a solution of a compound of the present invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid or benzoic acid. Many of these compounds of the invention carry an acidic group, in which case suitable pharmaceutically acceptable salts thereof may include alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), and formed salts. with suitable organic ligands such as quaternary ammonium salts. Also, in case an acidic (-COOH) or alcoholic group is present. pharmaceutically acceptable esters can be used to modify the solubility characteristics or hydrolysis of the compound.
The term administration and variants thereof (e.g., administration of a compound) in connection with a compound of the invention means providing the compound or prodrug of the compound to a subject in need of treatment. When a compound of the invention or a prodrug thereof is provided in combination with one or more other active agents (e.g., antiviral agents useful in the treatment of HCV infection), the use and variants thereof are understood to include the simultaneous and sequential provision of a compound or salt. (or hydrates) and other means.
As used herein, the term composition is defined to encompass a product containing said ingredients, as well as any product formed, directly or indirectly, by combining said ingredients.
Pharmaceutically acceptable means that the ingredients of the pharmaceutical composition must be mutually compatible and not harmful to the recipient.
The term subject (alternatively referred to herein as a patient) as used herein means an animal, preferably a mammal, most preferably a human, that has been subjected to treatment, observation or experiment.
The term effective amount as used herein means that amount of active compound or pharmaceutical agent that elicits a biological or medical response in a tissue, system, animal, or human that is required by a researcher, veterinarian, physician, or other physician. In one embodiment, the effective amount is a therapeutically effective amount for alleviating the symptoms of the disease or condition being treated. In another embodiment, the effective amount is a prophylactically effective amount for the prophylaxis of the symptoms of the disease or condition being prevented. The term also herein encompasses an amount of active compound sufficient to inhibit HCV NS3 protease and thereby elicit the desired response (i.e., an amount effective to
51101 Β inhibition). When the active compound (i.e., the active ingredient) is administered as a salt. reference to the amount of active ingredient refers to the form of the compound as the free acid or free base.
For the purpose of inhibiting HCV NS3 protease and preventing or treating HCV infection, the compounds of the present invention, optionally in the form of salts or hydrates, may be administered by any means that produce contact of the active agent with the site of action of the agent. They can be administered by any of the conventional agents available for use in conjunction with pharmaceutical agents, either as single therapeutic agents or in combination with therapeutic agents. They may be administered individually, but are typically administered with a pharmaceutical carrier selected based on the chosen route of administration and standard pharmaceutical practice. The compounds of the invention may, for example, be administered orally, parenterally (including subcutaneous injection techniques, intravenous, intramuscular, intrastemal injection or infusion), by inhalation spray, or rectally, in unit dosage form of a pharmaceutical composition containing an effective amount of the compound and conventional non-toxic pharmaceutically acceptable carriers, adjuvants and carriers. Liquid preparations suitable for oral administration (e.g., suspensions, syrups, elixirs, and the like) may be administered according to the techniques known in the art and may use any of the conventional methods such as water, glycols. oils, alcohols and the like. Solid preparations suitable for oral administration (e.g., powders, pills, capsules, and tablets) may be prepared according to techniques known in the art and may use such solid excipients as starches, sugars, kaolin, lubricants, binders. , disintegrants and the like. Parenteral compositions may be prepared according to techniques known in the art and typically use sterile water as a carrier and optionally other ingredients, such as a solubility enhancer. Injectable solutions may be prepared according to methods known in the art, wherein the carrier comprises saline, glucose solution or a solution comprising a mixture of saline and glucose. An additional description of methods suitable for use in the preparation of pharmaceutical compositions according to the present invention and ingredients suitable for use in said compositions is given in Remington's Pharmaceutical Sciences, 18th edition, edited by AR Gennaro, Mack Publishing Co., 1990.
The compounds of the present invention may be administered orally in a dose range of 0.001 to 1000 mg / kg of mammalian body weight (e.g., human) per day in a single dose or in divided doses. One preferred dose range is 0.01 to 500 mg / kg body weight per day in a single dose or in divided doses. Another preferred dose range is 0.1 to 100 mg / kg body weight per
51101 Β given orally in a single dose or in divided doses. For oral administration, the compositions may be presented as tablets or capsules containing 1.0 to 500 milligrams of active ingredient, in particular 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400 and 500 milligrams of active ingredient for symptomatic dose adjustment according to the patient being treated. The specific dose level and frequency of dosing for each patient may vary and will depend on various factors including the activity of the specific compound used, metabolic stability and duration of action of the compound, age, body weight, general health, sex, diet, route and time of administration. excretion rate. combination of drugs. the severity of the particular condition and the host undergoing therapy.
As stated above, the present invention also relates to a method for inhibiting HCV NS3 protease, inhibiting HCV replication, or preventing or treating HCV infection with a compound of the present invention in combination with one or more therapeutic agents and a pharmaceutical composition comprising the compound. according to the present invention and one or more therapeutic agents selected from the group consisting of HCV antiviral agent, immunomodulator and anti-infective agent. Such therapeutic agents that are active against HCV include, but are not limited to, ribavirin, levovirin, viramidine, thymosin alpha-1, R7025 (enhanced interferon (Roche)), interferon-β, interferon-α, pegylated interferon-α (peginterferon- a), a combination of interferon-α and fish A'irin, a combination of peginterferon-α and ribavirin, a combination of interferon-α and levovirin. and a combination of pcginterferon-α and levovirin. Interferon-α includes, but is not limited to. recombinant interferon-a2a (such as Roferon interferon available from Hoffmann-LaRochc, Nutlcy, NJ), pegylated interferon-a2a (Pegasys'<sup>M</sup>), interferon-a2b (such as lntron-A interferon available from Schering Corp., Kenilworth, NJ). pegylated interferon-α2b (Peglntron ™), recombinant consensus interferon (such as interferon alfacon-1), albuferon (interferon-α bound to human serum albumin (Human Genome Sciences)), and purified interferon-α product. Amgen's recombinant consensus interferon is protected by Infergen®. Levovirin is the L-enantiomer of ribavirin that has shown immunomodulatory activity similar to ribavirin. Viramidine is a ribavirin analogue listed in WO 01/60379 (assigned to ICN Pharmaceuticals). In accordance with the method of the present invention, the individual components of the combination may be administered separately at different times during therapy or simultaneously in divided or individual forms of combination.
For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with an agent that is an HCV NS3 inhibitor serine protease. HCV NS3 serine protease is an essential vinic enzyme and has been described as an excellent target site for
51101 Β inhibition of HCV replication. Substrate-based HCV NS3 protease inhibitors and non-substrate-based inhibitors are listed in WO 98/22496, WO 98/46630, WO 99/07733, WO 99/07734. WO 99/38888, WO 99/50230, WO 99/64442, WO 00/09543. WO 00/59929, GB-2337262, WO 02/48116, WO 02/48172 and U.S. Pat. 6,323,180.
Ribavirin, levovirin, and viramidine can exert their anti-HCV effects by modulating intracellular guanine nucleotide reserves through inhibition of the intracellular enzyme inosine monophosphate dehydrogenase (IMPJH). IMPDH is an enzyme that limits the rate of biosynthetic pathway in the de novo biosynthesis of guanine nucleotide. Ribavirin is easily phosphorylated intracellularly and the monophosphate derivative is an IMPDH inhibitor. Thus, inhibition of IMPDH represents the next useful target site for the discovery of HCV replication inhibitors. Accordingly, the compounds of the present invention may also be administered in combination with an IMPDH inhibitor, such as VX-497, which is disclosed in WO 97/41211 and WO 01/00622 (assigned to Vertex); another IMPDH inhibitor, such as that set forth in WO 00/25780 (assigned to Bristol-Myers Squibb); or mycophenolate mofetil [see AC Allison and EM Eugui, Agents Action, 44 (Suppl.): 165 (1993)].
For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with the antiviral agent amantadine (1aminoadamantane) [for a detailed description of this agent. see J. Kirschbaum, Anal. Profiles Drug Subs. 12: 1-36 (1983)].
For the treatment of HCV infection, the compounds of the present invention may also be administered in combination with an antiviral agent inhibiting R7128 (Roche) polymerase.
The compounds of the present invention may also be combined for the treatment of HCV infection with the antiviral 2'-C-garnet ribonucleosides listed in RE Harry-O'kuru, et al., J. Org. Chem., 62: 1754-1759 (1997); MS Wolfe, et al., Tetrahedron Lett., 36: 761 1-7614 (1995); US Pat. 3,480,613 (Nov. 25, 1969); International Publication No. WO 01/90121 (29 November 2001); International Publication Number WO 01/92282 (6 December 2001); and International Publication Number WO 02/32920 (April 25, 2002); and International Publication Number WO 04/002999 (8 January 2004); and International Publication Number WO 04/003000 (8 January 2004); and International Publication Number WO 04/002422 (January 8, 2004). Such 2'-c'-garnet ribonucleosides include, but are not limited to, 2'-C'-methyl-cytidine, 2 '- (' - methyl-uridine, 2'-C-methyl-adenosine. 2'-C-methyl-guanosine, and 9- (2-C-methyl-pD-ribofuranosyl) -2,6-diaminopurine, and the corresponding amino acid ester of ribose C-2 '. S-3 'and S-5' hydroxils and the corresponding optionally substituted cyclic 1,3-propanediol esters of 5'-phosphate derivatives.
51101 Β
The compounds of the present invention may also be combined for the treatment of HCV infection with other nucleosides that exhibit anti-HCV properties, such as those listed in WO 02/51425 (July 4, 2002), assigned to Mitsubishi Pharma Corp .; WO 01/79246, WO 02/32920, WO 02/48165 (June 20, 2002), and WO2005003147 (January 13, 2005) (including R1656, (2'A) -2'-deoxy-2'-fluoro-2 1-C-methikitidine, shown as compounds 3-6 on page 77) assigned to Pharmasset, Ltd .; WO 01/68663 (20.09.2001), assigned to ICN Phamiaceuticals; WO 99/43691 (2 September 1999); WO 02/18404 (7.03.2002), US2005 / 0038240 (17.02.2005) and WO2006021341 (2.03.2006), including 4'-azido nucleosides such as R1626, 4'azidocytidine, assigned to Hoffmann-LaRoche; US 2002/0019363 (14 February 2002); WO 02/100415 (19 December 2002); WO 03/026589 (3 April 2003); WO 03/026675 (3 April 2003); WO 03/093290 (13 November 2003); US 2003/0236216 (25.12.2003); US 2004/0006007 (January 8, 2004); WO 04/011478 (5 February 2004); WO 04/013300 (12 February 2004); US 2004/0063658 (April 1, 2004); and WO 04/028481 (8.04.2004).
For the treatment of HCV infection, the compounds of the present invention may also be used in combination with an agent that is an inhibitor of HCV NS5B polymerase. Such HCV NS5B polymerase inhibitors that can be used as combination therapy include, but are not limited to, those listed in WO 02/057287, US 6,777,395, WO 02/057425, US 2004/0067901, WO 03/068244. WO 2004/000858, WO 04/003138 and WO 2004/007512. Other such HCV polymerase inhibitors include, but are not limited to, valopicitabine (NM-283; Idenix) and 2'-F-2'-beta-methylcytidine (see also WO 2005/003147, assigned to Pharmasset, Ltd.).
In one embodiment, the nucleoside HCV NS5B polymerase inhibitors used in combination with the present HCV NS3 protease inhibitors are selected from the following compounds; 4-amino-7- (2-C-methyl-PD-arabinofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine: 4-amino-
7- (2-C-methyl-3-D-ribofuranosyl) -7H-pyrrolol2,3-d] pyrimidine; 4-methylamino-7- (2-C-methyl-β-dribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-dimethylamino-7- (2-C-methyl-PD-ribofuranosyl) 7H-pyrrolo [2,3-d] pyrimidine; 4-Cyclopropylamino-7- (2-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d (pyrimidine; 4-amino-7- (2-C-vinyl-PD-ribofuranosyl) -7H-pyrrolo [ 2,3-d] pyrimidine, 4 amino-7- (2-C-hydroxymethyl> PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine, 4-amino-7- (2-Cfluoromethyl-PD-ribofuranosyl) - 7H-pyrrolo [2,3-d] pyrimidine; 4-amino-5-methyl-7- (2-C-methyl-β-dribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-7- (2-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine-5-carboxylic acid; 4-amino-5-bromo-7- (2-C-methyl-p-Dribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-5-chloro-7- (2-C-methyl-n-ribofuranosyl) 7H-pyrrolo [2,3-d] pyrimidine; 4-Amino-5-fluoro-7- (2-C-phenyl-pD-ribofuranosyl) -7H-pyrrolo [2,321
51101 Β dpyrimidine; 2,4-diamino-7- (2-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 2 amino-7- (2-C-methyl-pD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 2-amino-4-cyclopropylamino-7- (2-C-methyl-p-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 2-amino-7- (2C-methyl-β-ribofuranosyl) -7H'pyrrolo [2,3-d] pyrimidin-4 (3H) -one; 4-amino-7- (2-C-ethyl-p-Dinbofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-7- (2-C, 2-O-dimethyl-p-ribofuranosyl) 7H-pyrrolo [2,3-d] pyrimidine; 7- (2-C-methyl-3-D-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidin-4 (3H) one; 2-amino-5-methyl-7- (2-C, 2-O-dimethyl-PD-ribofuranosyl) -7II-pyrrolo [2,3-d] pyrimidin4 (3H) -one; 4-amino-7- (3-deoxy-2-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-Carnino-7- (3-deoxy-2-C-methyl-PD-arabinofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-2-fluoro-7- (2-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-7- (3-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-7- (3-C-methyl-PD-xylofuranosyl) -711pyrrolo [2,3-d] pyrimidine; 4-amino-7- (2,4-di-C-methyl-PD-ribofuranosyl) -7H-pyrrolo [2,3-d] pyrimidine; 4-amino-7- (3-deoxy-3-fluoro-2-C-methyl-p-ribofuranosyl) -71H-pyrrolo [2,3-d] pyrimidine; and the corresponding 5'-triphosphates; or a pharmaceutically acceptable salt thereof.
The compounds of the present invention may also be combined for the treatment of HCV infection with non-nucleoside IICV polymerase inhibitors such as those disclosed in WO 01/77091 (18.10.2001), assigned to Tularik, Inc .; WO 01/47883 (July 5, 2001), assigned to Japan Tobacco, Inc .; WO 02/04425 (17 January 2002), assigned to Boehringer Ingelheim; WO 02/06246 (24 January 2002), awarded by the Istituto di Ricerche di Biologia Moleculare P. Angeletti
SPA; WO 02/20497 (March 3, 2002); WO 2005/016927 (in particular JTK003), assigned to Japan Tobacco, Inc .; and HCV-796 (Viropharma Inc.).
In one embodiment, the non-nucleoside HCV NS5B polymerase inhibitors used in combination with the present HCV NS3 protease inhibitors are selected from the following compounds: 14-cyclohexyl-6- [2- (dimethylamino) ethyl] -7-oxo-5,6 , 7,8-tetrahydroindolo [2.1a] [2.5] benzodiazocine-1-carboxylic acid; 14-cyclohexyl-6- (2-morpholin-4-ylethyl) -5,6,7,8-tetrahydroindolo [2,1] [2,5] benzodiazocine-1-carboxylic acid; 14-cyclohexyl-6- [2- (dimethyl amino) ethyl] -3-methoxy-5,6,7,8-tetrahydroimdolo [2,1-f] [2,5] benzodiazole-11-carboxylic acid; 14-cyclohexyl-3-methoxy-6-methyl-5,6,7,8-tetrahydroindolo [2,1a] [2,5] benzodiazocine-11-carboxylic acid; inethyl ({[(14-cyclohexyl-3-methoxy-6-methyl-5,6,7,8-tetrahydroindolo [2.1-a] [2.5] benzodiazocin-1-yl) carbonyl] amino} sulfonyl) acetate; ({[(14-cyclohexyl-3-methoxy-
6-methyl-5,6,7,8-tetrahydroindolo [2,1a] [2,5] benzodiazocin-11-yl) carbonyl] amino] sulfonyl) acetic acid; 14-cyclohexyl - N - [(dimethylamino) sulfonyl] -3-methoxy-6-methyl-5,6,7,8-tetrahydroindolo [2,1-f] [2,5] benzodiazocin-1-carboxamide: 3-chloro -14-cyclohexyl-6- [2- (dimethylamino) ethyl] -7-oxo-5,6,7,8-tetrahydroindolo [2.1-a] [2,5] benzodiazocin 11-carbon
51101 Β acid; N- (1-Carboxy-1,4-cyclohexyl-7,8-dihydro-6H-indolo [1,2-e] [1,5] benzoxazocin-7-yl).<sup>r</sup>-dimethylelane-1,2-diaminium bis (trifluoroacetate); 14-cyclohexyl-7,8-dihydro-6H-indolo [1,2-e] [1,5] benzoxazocine-1-carboxylic acid; 14-cyclohexyl-6-methyl-7-oxo-5,6,7,8-tetrahydroindolo [2,1-c] [2,5] benzodiazocin-1,1-carboxylic acid; 14-cyclohexyl-3-methoxy-6-methyl-7-oxo-5,6,7,8-tetrahydroindolo [2,1] [2,5] benzodiazocin-11-carboxylic acid; 14-cyclohexyl-6- [2- (dimethylamino) ethyl] -3-methoxy-7-oxo-5,6,7,8-tetrahydroindolo [2,1a] [2,5] benzodiazocine-11-carboxylic acid; 14-cyclohexyl-6- [3- (dimethylamino) propyl] -7-oxo5,6,7,8-tetrahydromido [2,1] [2,5] benzodiazole-1-carboxylic acid; 14-cyclohexyl-7-oxo-6- (2-piperidin-1-ylethyl) -5,6,7,8-tetrahydroindolo [2,1] [2,5] benzodiazocin-1-carboxylic acid; 14-cyclohexyl-6- (2-morpholin-4-ylethyl) -7-oxo-5,6,7,8-tetrahydroindolo [2,1] [2,5] benzodiazocin-11-carboxylic acid; 14-cyclohexyl-6- [2- (diethylamino) ethyl] -7-oxo-5,6,7,8-tetrahydroindolo [2,1-b] [2,5] benzodiazocin-1-carboxylic acid; 14-cyclohexyl-6- (1-methyl piperidin-4-yl) -7-oxo-5,6,7,8-tetrahydroindolo [2,1-f] [2,5] benzodiazocin-11-carboxylic acid; 14-cyclohexyl-N - [(dimethylamino) sulfonyl] -7-oxo-6- (2-piperidin-1-ylethyl) -5,6,7,8tetrahydroindolo [2,1] [2,5] benzodiazocin-11- carboxamide; 14-cyclohexyl-6- [2- (dimethyl amino) ethyl] -<sub>.</sub>N - [(dimethylamino) sulfonyl] -7-oxo-5,6,7,8-tetrahydroindolo [2,1-b] [2,5] benzo diazocin-11-carboxamide; 14-cyclopentyl-6- [2- (dimethylamino) ethyl] -7-oxo-5,6,7,8-tetrahydroindolo [2,1-b] [2.5] benzodiazocin-1-carboxylic acid; 14-cyclohexyl-5,6,7,8tetrahydroindolo [2,1a] [2,5] benzodiazocine-1-carboxylic acid; 6-allyl-14-cyclohexyl-3-methoxy-5,6,7,8-tetrahydroindolo [2.1-a] [2,5] benzodiazocin-1-carboxylic acid; 14-cyclopentyl] -6- [2- (dimethylamino) ethyl] -5,6,7,8-tetrahydroindolo [2,1-f] [2,5] benzodiazocin-1-carboxylic acid; 14-cyclohexyl-6- [2- (dimethylamino) ethyl] -5,6,7,8-tetrahydropyridolo [2,1] [2,5] benzodiazocin-1-carboxylic acid; 13-cyclohexyl-5-methyl-4,5,6,7-tetrahydrofuro [3 ', 2': 6,7] [1,4] diazocino [1,8-yl] indole-10-carboxylic acid; 15-cyclohexyl-6- [2 (dimethylamino) ethyl] -7-oxo-6,7,8,9-telrahydro-5H-indolo [2,1-yl] [2,6] benzodiazonine-12-carboxylic acid; 15-Cyclohexyl-8-oxo-6,7,8,9-letrahydro-57H-indolo [2.1 i] [2,5] benzodiazonine-1,2-carboxylic acid; 13-cyclohexyl-6-oxo-6,7-dihydro-5H-indolo [1,2-yl] [1,4] benzodiazepine-10-carboxylic acid; and pharmaceutically acceptable salts thereof.
The aforementioned indole-based tetracyclic HCV NS5B polymerase inhibitors can be obtained by monitoring AE methods as described below, where different variables can be selected according to the specific tetracyclic indole compound to be prepared:
51101Β
Procedure Α
<img file="RS51101B_D0029.tif" />
<img file="RS51101B_D0030.tif" />
<img file="RS51101B_D0031.tif" />
“PdU '
<img file="RS51101B_D0032.tif" />
deprotection
-<-------------- ' '
<img file="RS51101B_D0033.tif" />
The 2-bromoindole intermediate (prepared as described in published International Patent Application WO2004087714) was functionalized on indole nitrogen to introduce the precursor functional group W7X 'on either or both elements of the W / X chain. The Pd-mediated cross-coupling methodology (e.g., Suzuki, Stille z7z /.) Was then introduced into the precursor functional group Ζ7Υ 'bearing a C2 aromatic ring, on either or both elements of the Ζ / Υ chain. Manipulation of the functional group followed by ring closure gave a tetracyclic system. Deprotection of the ester then gave the target carboxylic acid indoles. with a C2 aromatic ring attached to the indole nitrogen.
Procedure B
<img file="RS51101B_D0034.tif" />
A
X'-W'-halogen
<img file="RS51101B_D0035.tif" />
closing 'the vertebrae
L w
H / Br
1) manipulation of functional gnipotn
2) RO formation<sub>2</sub>C
<img file="RS51101B_D0036.tif" />
<img file="RS51101B_D0037.tif" />
<img file="RS51101B_D0038.tif" />
<img file="RS51101B_D0039.tif" />
51101Β
After chain formation outside the corresponding 2-haloaromatic ring, the mediated ring closure gave a fused tetracyclic system. Deprotection of the ester then gave the target indole carboxylic acid, with the C2 aromatic ring attached to the indole nitrogen.
Procedure C
<img file="RS51101B_D0040.tif" />
<img file="RS51101B_D0041.tif" />
The C2 aromatic ring was introduced as indicated at the beginning via a Pd-mediated cross-linking methodology (Suzuki, Stille, etc.). A chain was then formed, cyclizing to the indole nitrogen finally closing the ring. Deprotection of the ester then gave the target indole carboxylic acids , with a C2 aromatic ring attached to the indole nitrogen.
Procedure D
<img file="RS51101B_D0042.tif" />
1) manipulation of functional gnipoill
2) deprotection
<img file="RS51101B_D0043.tif" />
The fused tetracyclic intermediates resulting from Methods AC are subjected to manipulation of the functional group in the chain prior to ester deprotection to obtain the target C2-linked indole carboxylic acids.
51101Β
Procedure Ε
<img file="RS51101B_D0044.tif" />
The C2-linked indole carboxylic acids obtained by Method AD were further derivatized by manipulating the carboxylate functional group to give compounds carrying a carboxylate substitution or carboxamide. During any of the aforementioned synthetic methods, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules in question. This can be accomplished using conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & PGM Wuts, Protective Groups in Organic Synthcsis. John Wiley & Sons, 3rd edition. 1999. The protecting groups can be removed at an appropriate later stage using methods known in the art.
The inhibitory activity of the present compounds against HCV NS3 protease can be tested using assays known in the art. One such assay is the time-decomposed fluorescent (TRF) IICV NS3 protease assay as described in Example 56. Other examples of such assays are described in, e.g., International Patent Publication WO2005 / 046712. Compounds useful as HCV NS3 protease inhibitors would have a Ki of less than 50 μΜ, more preferably less than 10 μΜ, and even more preferably less than 100 nM.
The present invention also encompasses methods for preparing compounds of formula I, II or III. The compounds of the present invention can be readily prepared according to the following reaction schemes and examples, or modifications thereof, using readily available starting materials, reagents and conventional synthetic methods. In these reactions, it is also possible to use variants which are known per se to those skilled in the art, but are not described in more detail. In addition, other methods for preparing the compounds of the invention will be readily apparent to those skilled in the art in light of the following reaction schemes and examples. Unless otherwise indicated. All variables are as defined above. The following reaction schemes and the examples serve only to illustrate the invention and its implementation.The examples should not be construed as limiting the scope and meaning of the invention.
51101 Β
General description of the synthesis:
The compounds of the present invention may be synthesized as set forth in General Schemes 1 and 2.
ШЕМАл (Кđ1.2 (Р<sup>5</sup>)! - 2) l, 2
1) Removal <mje Boc
2) Amide coupling
--— ј ».
<img file="RS51101B_D0045.tif" />
1) optional hydrogenation or functionalization
2) Estarska hi '
3) K.amide influx) l, 2> 0
Metathesis) 1,2> 0
Scheme 1 (n = 0-9) shows the synthesis of a representative molecule. A suitably protected 4-hydroxyproline derivative (e.g., carbonate-protected nitrogen and ester-protected acid) can be reacted with carbonyldiimidazole or an equivalent reagent and then reacted with an appropriately substituted isoindoline or tetrahydroisoquinoin. The alkenyl functional group can be introduced at this or a later stage by a reaction which is catalyzed by a palladium halide substituent such as chloride, bromide and iodide, or another functional group such as a triflate with an organometallic reagent such as vinyl or allyltrialkyltin. Alternatively, the alkenyl functional group may be introduced prior to reaction with the protected prolinol.
Scheme 2 describes the synthesis of an olefin-containing amino acid moiety. An amino acid (either commercially available or can be readily prepared using methods known in the art) in which the acidic functional group is protected as an ester (for example, R-melyl) can be converted to amides A by coupling olefinic carboxylic acid using a broad
51101Β ranges of peptide coupling agents known to those skilled in the art such as DCC, EDC, BOP, TBTU, etc. Preparation of sulfonamide B can be accomplished by reaction with the appropriate sulfonyl chloride in an organic solvent (e.g., THF) with an amine base as a scavenger. Urea C derivatives can be prepared by reacting the amino ester with a reagent such as carbonyldiimidazole to form an intermediate isocyanate (Catalano et al., WO 03/062192) followed by the addition of another olefin-containing amine. Alternatively, phosgene, diphosgene or triphosgene may be used instead of carbonyldiimidazole. Cyanoguanidine derivatives D can be prepared by reacting an amino acid ester with diphenyl C-cyanocarbonimidate in an organic solvent. followed by the addition of another olefin-containing amine. Carbamate E derivatives can be prepared by reacting an olefin-containing alcohol with carbonyldiimidazole (or phosgene, triphosgene or diphosgene) in an organic solvent. followed by the addition of an amino ester.
SCHEME2
<img file="RS51101B_D0046.tif" />
After functionalization of the amine, the ester can be hydrolyzed under a range of basic conditions known to those skilled in the art (Theodora W. Grccnc, Protective Groups in Organic Synthesis, Third Edition. John Wiley and Sons, 1999).
Deprotection of the carbamate protecting group on the proline portion can be performed using various methods known to those skilled in the art (Theodora W. Greene, Protective Groups in Organic Synthesis, Third Edition, John Wiley and Sons. 1999).
51101 Β
To complete the synthesis of the compounds of the present invention, an amino acid derivative can be coupled to a proline derivative using a wide range of peptide coupling reagents such as DCC, EDC, BOP, TBTU and the like. (see Scheme 1). Macrocyclization is then achieved via olefin metathesis using a range of catalysts described in the literature for this purpose. At this stage, the olefinic bond produced in the ring closure metathesis can be optionally hydrogenated to obtain a saturated bond or functionalized in alternative ways such as cyclopropanation. The proline ester is then hydrolyzed under basic conditions and combined with cyclopropylamino acid ester (the corresponding alkenyl or alkylcyclopropane portion of the molecule can be prepared as previously described) (Llinas-Brunet et al., US 6,323,180) and subjected to an additional basic hydrolysis step to provide final compounds. The proline ester can also be hydrolyzed and directly coupled with appropriately functionalized cyclopropylamino acid acyl sulfonamide (which can be prepared according to Wang H.A. et al. WQ2003 / 099274) to provide the final compounds.
Catalysts for olefin metathesis include the following ruthenium-based species: F: Miller et al J. Am. Chem. Soc 1996, 118, 9606; G: Kingsbury et al J.Am. Chem. Soc 1999, 121, 791; H: Scholl et al Org. Lett. 1999, 1, 953; Hovcyda et al US2002 / 0107138; K: Furstner et al. .1. Org. Chem 1999, 64, 8275. The benefit of these catalysts in ring closure metathesis is well known in the literature (e.g., Tmka and Grubbs, Acc. Chem. Res. 2001, 34, 18).
<img file="RS51101B_D0047.tif" />
<img file="RS51101B_D0048.tif" />
<img file="RS51101B_D0049.tif" />
<img file="RS51101B_D0050.tif" />
<img file="RS51101B_D0051.tif" />
<img file="RS51101B_D0052.tif" />
Zhan Iutenium Melatese Catalyst RC-303 (Zhan CatalystlB, RC-303, Zannan Phanna Ltd.)
51101 Β
List of abbreviations
BOP Benzotriazol-1-yl-oxy-tris- (dimethylamino) -phosphonium hexafluorophosphate
DCC. Dicyclohexylcarbodiimide CH3CN Acetonitrile
<td>DBU</td><td>1,8-Diazabicyclo [5.4.0] undec-7-ene</td><td></td><td></td>
<td>DCE</td><td>Dichloroethane</td><td>DCM</td><td>Dichloromethane</td>
<td>DMAP</td><td>4-Dimethylamino pyridine</td><td>DIPEA</td><td>Diisopropylethylamine</td>
<td>DMF</td><td>Dimethylformamide</td><td>ЂМС (^</td><td>Dimethyl sulfoxide</td>
<td>EDC</td><td colspan="2">N- (3-Dimethylaminopropyl) -N-ethylcarbodiimide</td><td></td>
<td>Et<sub>3</sub>N</td><td>Triethylamine</td><td>EbO</td><td>Diethyl ether</td>
<td>EtOAc</td><td>: Ethyl acetate</td><td>EtOH</td><td>Ethanol</td>
<td>HATU</td><td>O- (7-Azabenzotriazol-1-yl) -AEV, A</td><td colspan="2">B'-tetramethyluronium hexafluorophosphate</td>
<td>HBr</td><td>Hydrobromic acid</td><td></td><td></td>
<td>HCl</td><td>Hydrochloric acid</td><td>HOAc</td><td>Acetic acid</td>
<td>HOAt</td><td>1-hydroxy-7-azabenzotriazole</td><td>I.iOH</td><td>Lithium hydroxide</td>
<td>MeOH</td><td>Methanol</td><td>MgSO<sub>4</sub></td><td>Magnesium sulfate</td>
<td>NaHCO<sub>3</sub></td><td>Sodium bicarbonate</td><td>On<sub>2</sub>SO<sub>4</sub></td><td>Sodium sulfate</td>
<td>NaOH</td><td>Sodium hydroxide</td><td>, NH<sub>4</sub>C1</td><td>Ammonium chloride</td>
<td>nh<sub>4</sub>oh</td><td>Ammonium hydroxide</td><td>Pd <:</td><td>Palladium on coal</td>
Pdi'PPhjj .tetrakis (triphenylphosphii) palladium (0)
<td>PhMc</td><td>Toluene</td><td>PPh<sub>5</sub></td><td>Triphenylphosphine</td>
<td>RT</td><td>Room temperature</td><td>ΑΉΡ</td><td>Tetrahydofuran</td>
TBTU O-Benzotriazol-1-yl-N, N, N Ά-tetramethyluronium tetrayl fluoroborate
51101 Β
EXAMPLE 1 (5R, 7S, 10S) -10-Butyl-N - ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropyl) -3,9,12-trioxo-1,6 7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8dimethano-4,13,2,8,11-benzothioxatriazacycloicosine-7-carboxamide (III-1)
<img file="RS51101B_D0053.tif" />
Step 1: 4-Chloroisoindoline
<img file="RS51101B_D0054.tif" />
A mixture of 3-chlorophthalic acid anhydride (9 g, 49.2 mmol) and formamide (100 mL) was heated to 125 ° C and stirred for 3 hours. Water (300 mL) was added and the mixture was cooled to room temperature. The mixture was filtered and the resulting white solid was washed with water and dried to give 4-chloro-1H-isoindole-L3 (2H) -dione (7.7 g, 86% yield).
To the solid 4-chloro-1H-isoindole-1,3 (2H) -dione (4.0 g, 22.0 mmol) was added dropwise the borane-THF complex (1 M / THF, 88.1 mL, 88.1 mmol) with stirring. When the addition was complete, the reaction mixture was heated to reflux (80 ° C)<sup>o</sup>C) and stirred for 6 hours. The reaction mixture was then cooled to 0 ° C, methanol (2.8 mL, 88.1 mmol) was carefully added dropwise and the reaction mixture was warmed to room temperature. HCl (6 N) was added until the mixture became acidic and then the mixture was concentrated. The crude product was dissolved in 1 M HCl and extracted twice with ethyl ether and twice with dichloromethane. The pH of the aqueous layer was adjusted to pH = 11 with solid NaOH and it was extracted three times with ethyl acetate. The combined ethyl acetate extracts were dried rgeko Na<sub>2</sub>SO4, filtered and
51101Β concentrated to give 4-chloroisoindoline (1.8 g, 53% yield). LRMS (ESI) m / z 154 [(M + H)<sup>+</sup>; calc. zaC<sub>g</sub>H<sub>9</sub>ClN: 154].
Step 2: 1-tert-Butyl 2-methyl (2S, 4R) -4-B (4-chloro-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate
<img file="RS51101B_D0055.tif" />
To a solution of N-Boc proline methyl ester (2.87 g, 11.7 mmol) in DMF (15 mL) at 0 ° C was added carbonyldiimidazole (1.9 g, 11.7 mmol). The reaction was warmed to room temperature and stirred for 30 min. A solution of 4-chloroisoindoline (1.8 g, 11.7 mmol) in DMF (10 mL) was then added and the reaction mixture was heated to 50 ° C and stirred for 2 hours. The reaction mixture was poured onto ethyl ether and 0.5 M HCl, and the layers were separated. The organic layer was washed with water, dried over Na<sub>2</sub>SO<sub>4</sub>, filtered and concentrated. The crude product was purified on silica gel (eluting with a gradient of 10% to 90% ethyl acetate in hexanes) to give 1-to-c-butyl 2-methyl (2S, 4R) -4 - {[(4-chloro- 1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate (3.3 g, 66% yield). LRMS (ESI) m / z 325 [(M + H-Boc)<sup>+</sup>; calc. for C<sub>15</sub>H<sub>18</sub>C1N<sub>2</sub>Oh<sub>4</sub>:325],
Step 3: 1-tert-Butyl 2-methyl (2S, 4R) -4 - {[(4-vinyl-13-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate
<img file="RS51101B_D0056.tif" />
51101 Β
1-tert-butyl 2-methyl (2S.4R) -4 - [[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate solution (40 mg , 0.09 mmol), vinyl tributylstannane (36 mg, 0.11 mmol) and cesium fluoride (31 mg, 0.21 mmol) in dioxane (0.5 mL) were degassed with N<sub>2</sub> for 15 min. Bis (tributylphosphine) palladium (0) (2 nig, 0.005 mmol) was then added and the reaction vessel was sealed and heated to 100 ° C for 18 hours. After cooling, the reaction mixture was concentrated and purified by chromatography on silica gel (10% to 90% ethyl acetate in hexanes) to give 1-tert-butyl 2-methyl (2S, 4R) -4 - ([(4vinyl- 1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate (10 mg, 25% yield) LRMS (ESI) m -1 317 [(M + II-Boc)<sup>+</sup>: calc. for Ci<sub>7</sub>H<sub>21</sub>N<sub>2</sub>Oh<sub>4</sub>: 317],
Step 4: Methyl N - [(pent-4-enyloxy) carbonyl] -L-norylcyl- (4R) - {[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonylloxy} -L -proline
<img file="RS51101B_D0057.tif" />
In a vessel containing 1-zerc-butyl 2-methyl (2S.4R) -4 - {[(4-vinyl-1,3-dihydro-2Hisoindol-2-yl) carbonyl] oxy) pyrrolidine-1,2-dicarboxylate mg, 0.14 mmol) was added to a 4 M solution of HCl in dioxane (2 mL). After 1 hour, LC-MS analysis showed complete consumption of starting material and formation of the desired Boc product. The volatiles were then removed in vacuo, and the crude material was poured into DMF (2 mL).
To this mixture was added A4 (pent4-en-1-yloxy) carbonyl] -L-norleucine (41 mg, 0.17 mmol) (prepared according to the procedure below), DIPEA (0.076 mL. 0.43 mmol), EDC (54 mg, 0.28 mmol) and HOAt (44 mg, 0.28 mmol). After stirring at room temperature for 30 min. complete amine consumption was demonstrated via LC-MS. The reaction mixture was then treated with 0.5 N HCl and EtOAc. The organic layer was washed with brine and dried over MgSO 4<sub>4</sub>. The solvent was then removed in vacuo and the crude product was purified on silica (10-90% EtOAc / hexanes) to give 60 mg (79% yield) of methyl A<sup>r</sup>- [(pent-4-enyloxy) carbonyl] -L-norleucyl- (4R) -4-t [(4-vinyl-1,3-dihydro-2-isoisolindol-2-yl) carbonyl] oxy} -L-prolineate. LRMS (ESI) m + 542 [(M + H) + calcd. for CboFLfoNjO ;: 542],
51101Β
Step 5: Methyl (5R, 7S, 10S) -10-butyl-3,9,12-trioxo-1,6,7,9,10,11,11,14,15,16dehydro-5H-2,22: 5,8-dimethatio-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxylal
<img file="RS51101B_D0058.tif" />
Methyl A - [(pent-4-enyloxy) carbonyl] -L-norleucyl- (4R) -4 - {[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} -L solution -proline (60 mg, 0.11 mmol) in DCE (20 mL) was degassed with N? for 15 min. Then Zhan ruthenium metathesis catalyst RC-301 (Zhan catalyst I (shown as J on page 43), RC-301, Zannan Pharma Ltd.) (7 mg, 0.01 mmol) was added. The solution was then heated to 100 ° C for 1 hour. At that time, LC-MS and TLC analysis showed complete consumption of starting material and formation of an almost single product having the desired mass. The solvent was then removed in vacuo. and crude products purified on silica (5-70% EtOAc / hexane) to give 45 mg (79% yield) of methyl (5R, 7S, 10S) -10-butyl-3,9,12-trioxo-1, 6,7,9,10,11,1,1,14,15.16-decahydro-5H-2,22: 5,8dimethano-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxylate. LRMS (ESI) m + 514 [(M + H)<sup>+</sup>; calc. for C77H36N3O7: 514].
Step 6: (5R, 7S, 10S) -10-Butyl-N - ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl] -2-vinylcyclopropyl) -3,9,12-trioxo-1,6) , 7,9,10,11,1,1,14,15,16-decahydro-5H2,22: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxanide
To a solution of methyl (5R, 7S, 10S) -10-butyl-3,9,12-trioxo-1,6,7,9.10,11,1,1,14,15,16decahydro-5H-2,22: 5, 8 -dimethano-4,13,2,8,1 l-benzodioxatriazacycloicine-7-carboxylate (45 mg, 0.09 mmol) in TIIF (2 mL), MeOH (0.5 mL) and water (1 mL) was added LiOH (21 mg) , 0.87 mmol). The reaction mixture was heated to 40 ° C and stirred for 1 hour, after which complete consumption of methyl ester starting material was recorded by LC-MS. The mixture was then treated with 0.5 N HCl and EtOAc. The organic layer was then dried over K2CO3, and the solvent was removed in vacuo. The crude product was poured into DMF '(I mL).
To the above solution was added (1R, 2S) -1 -] [(cyclopropylsulfonyl) aminocarbonyl} -2-vinylcyclopylopanaminium chloride (IJinas-Brunet et al. US03 / 15755 and Wang et al.
51101 Β
WO 03/099274) (32 mg, 0.12 mmol), TBTU (51 mg, 0.16 mmol) and DIPEA (0.071 mL, 0.40 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was directly purified by reverse phase IIPLC to give (5R, 7S, 10S) -10-butyl-A ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropyl) - 3,9,12-trioxo1,6,7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8-dimethano-4,13,2,8,11benzodioxatriazacycloicosin-7- carboxamide (27 mg. 47% yield). 1 H NMR (500 MHz, ppm. CDCl 3)<sub>3</sub>) b 10.01 (s, 1H), 7.27 (m, 2H), 7.12 (d, 1H). 7.04 (s, 1H). 6.40 (d, J = 16.1 Hz, 1H), 6.08 (m, 1H), 5.76 (m, 1H), 5.44 (s, 1H), 5.36 (d, 1H), 5.25 (d, 1). H), 5.14 (d. 1H), 4.804.68 (m, 3H). 4.59 (d, 1H), 4.44 (m, 2H). 4.38 (m, 1H), 4.28 (m, 1H). 3.95 (m, 1H), 3.77 (dd, 1H), 2.94 (m, 1H), 2.43 (m, 2H), 2.29 (d, 2H), 2.06 (m<sub>:</sub> 2 Η), 1.94 (m, 1 Η), 1.78 (m, 4 H). 1.45 (m, 1H), 1.38-1.06 (m, 5H), 1.04 (d, 2H), 0.92 (t, 3H) ppm. LRMS (ESI) 712 (M + H)<sup>+</sup>; calc. for C35H46N5O9S: 712J.
EXAMPLE 2 (5R, 7S, 10S) -10-tert-Butyl-X - ((1R, 2S) -1-ii-cyclopropylsulfonyl) aminocarbonyl} -2-vinylcyclopropyl) -3,9,12-trioxo-1,6,7 9,10,11,1,1,14,15,16-decahydro-5H-2,22: 5Sdimethano-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxamide (11-2)
<img file="RS51101B_D0059.tif" />
EXAMPLE 2 was prepared according to the procedure used for EXAMPLE 1 with the difference that 3-methyl-N - [(pent-4-enyloxy) carbonyl] -L-valine (prepared according to the procedure below) was used instead N- (pent-4-en-1-yloxy) carbonyl] -L-norleucine in Step 4. 1 N NMR (500 MHz, ppm, CDCl 3)<sub>3</sub>) b 9.90 (s, 1H), 7.28 (m, 2H), 7.13 (m, 2 II), 6.31 (d, .7 = 15.9 Hz, 1H). 6.04 (m, 1H). 5.74 (m, 1 H), 5.45 (m, 2 H), 5.27 (d, 1 II), 5.16 (d, 1 H), 4.77-4.66 (m, 3 H), 4.55 (d, 1 H), 4.48 (t, 1H). 4.41-4.35 (m, 2H), 4.27 (m, 1H), 3.93 (m, 1H), 3.74 (dd, 1H), 2.93 (m, 1H), 2.45 (d, 2H). 2.32 (m, 2H). 2.10-1.95 (m, 2H). 1.74
51101Β (m, 1 Η), 1.47 (m, 1 Η), 1.37 (m. 2 Η), 1.07 (p. 9 Η) ppm. LRMS (ESI) m / z 712 [(M + H +; calcd. GaSzbNdbMzOcZ: 712].
EXAMPLE 3 (5R, 7S, 10S) -10- [erc-Boutique N- ((1R, 2S) -1- {Cyc] propropylsulfonylamino] carbonyl) -2vinylcyldopropyl) -15,15-dimethyl-3,9,12-trioxo -l<sub>1</sub>6,7,9,10,11,12,14,15,16-decahydro-5H2,22: 5,8-dimethano-4H3,2,8,11 benzodioxatriazacycloicosine-7-carboxamide (W-8)
<img file="RS51101B_D0060.tif" />
Step 1: 1-Bromo-2 3-bis (bromomethyl) benzene
Br
<img file="RS51101B_D0061.tif" />
Br Vg
3-bromo-o-xylene suspension (196 g, 1.06 mol). N-bromosuccinimide (377 g, 2.15 mol) and benzoyl peroxide (0.26 g, 1.0 mmol) in carbon tetrachloride (1800 mL) were heated to reflux under nitrogen for 15 hours. The contents of the reaction vessel were cooled. filtered and the filtrate was evaporated. The crude material was distilled under high vacuum. The main fractions were determined between 88 ° C and 152 ° C. 108 g of pure material are obtained. 182 g of slightly crude material are obtained which can be used in the next reaction. 1 H NMR (CDCl 3)<sub>3</sub>) b (ppm) 7.56 (d, J = 8.0 Hz, 1 H), 7.31 (d, J = 8.0 Hz)<sub>?</sub> 1H), 7.26 (s, 1H), 7.16 (t, J = 8.0 Hz, 1H), 4.84 (s, 2H). 4.64 (s, 2H).
51101 Β
Step 2: 2-Benzyl-4-bromoisomidoline
<img file="RS51101B_D0062.tif" />
Potassium bicarbonate (204 g, 2.04 mol) was suspended in acetonitrile (12 L) and the mixture was heated to 80 ° C. Solutions of 1-bromo-2,3-bis (bromomethyl) benzene (280 g, 0.82 mol in 500 mL acetonitrile) and benzylamine (87.5 g, 0.82 mol in 500 mL acetonitrile) were added simultaneously via addition funnel over 1 hour. The reaction mixture was stirred at 77 ° C for 16 hours. The contents of the reaction vessel were cooled, filtered and the solvent was removed by evaporation. The reaction was partitioned between 1M K<sub>2</sub>CO<sub>3</sub> and EtOAc. The organic niateria were washed with brine, dried over anhydrous Na<sub>2</sub>SO4, filtered and evaporated. Flash chromatography (gradient elution: heptane to 10% EtOAc in heptane) after evaporation gave the title compound as a pale oil. 11 NMR (CDCl 3)<sub>3</sub>) b (ppm) 7.41-7.39 (m, 2H), 7.37-7.34 (m, 2H), 7.32-7.27 (m, 2H), 7.10-7.03 (m, 2H), 4.02 (s. 2 H), 3.97 (s, 2H). 3.91 (s, 2H). LRMS (ESI) m / z 289 [(M + H & lt; + & gt;)<sup>+</sup>; calc. for C |<sub>5</sub>H<sub>15</sub>BrN: 289],
Conversion to HCl salt to HCl / MeOH was performed. MTBE was added and the solid was filtered to give 118 g of product as the HCl salt.
Step 3: 2-Benzyl-4-vinylisoindoline
<img file="RS51101B_D0063.tif" />
A solution of 2-benzyl-4-bromoisoindoline (16.7 g, 58.0 mmol) and tributyl (vinyl) tin (20.3 mL, 69.6 mmol) in toluene (400 mL) was degassed by passing nitrogen gas through the solution for 0.25 h. Tetrakis (triphenylphosphine) palladium (0) (1.30 g, 1.16 mmol) was added and the resulting solution was heated in an oil bath at 100 ° C under nitrogen for 24 hours. The contents of the reaction vessel were cooled, evaporated and subjected to flash column chromatography eluting with hexane / ethyl acetate 95/5 to give, after evaporation, the title compound as a pale oil which turned pink on standing. LRMS (ESI) m / z 236 [(M + H)<sup>+</sup>; calc. for C<sub>I7</sub>H<sub>1;(</sub>N: 236],
51101Β
Step 4: 4-Vinylisoindoin
<img file="RS51101B_D0064.tif" />
A solution of 2-benzyl-4-vinylisoindoline (58 mmol) in 1,2-dichloroethane (150 mL) was placed in a 1L round bottom flask under nitrogen. To this was added a addition funnel containing a solution of 1-chloroethyl chloroformate (7.5.1 mL, 69.6 mmol) in 1,2-dichloroethane. The reaction vessel was cooled in an ice bath and the contents of the addition funnel were added dropwise over 20 min while maintaining the internal reaction temperature <5 ° C. After the addition was complete, the reaction vessel was allowed to warm to room temperature and then heated to reflux for 45 min. The contents of the reaction vessel were cooled to room temperature and then the solvent was removed by evaporation. Methanol (200 mL) was added and the contents of the reaction vessel were heated to reflux for 30 min. The reaction vessel was cooled and the solvent was removed by evaporation. Water (200 mL) was added and the resulting mixture was washed with ethyl acetate (2 x 250 mL). The aqueous layer was basified with 2N sodium hydroxide and then extracted with methylene chloride (4 χ 250 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and the filtrate was evaporated. The rest underwent ..flash '<sup>4 </sup>column chromatography eluting with methylene chloride / methanol / ammonium hydroxide 97/3 / 0.3 to 95/5 / 0.5. Evaporation of the fractions gave the title compound as a brown oil, 6.00 g (41.4 mmol, 71% yield in two steps). LRMS (ESI) m / z 146 calcd. for
C<sub>10</sub>H<sub>I2</sub>N: 146].
Step 5; 1-tert-Butyl 2-methyl (25,47?) - 4 - {[(4-vinyl-1,3-dihydro-2fl<sup>r</sup>-isoindol-2yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate
<img file="RS51101B_D0065.tif" />
Solution of 1-tert-butyl 2-methyl (25,4 / b) -4-hydroxypyrrolidine-1,2-dicarboxylate (10.1 g,
41.4 mmol) in DMF (90 mL) under nitrogen was cooled to 0 ° C. Solid 1.Γ was added to the reaction
51101Β
<img file="RS51101B_D0066.tif" />
carbonyldiimidazole (6.70 g, 41.4 mmol). The contents of the reaction vessel were warmed to room temperature and after 2 hours a solution of 4-vinylisoindoline (6.00 g, 41.4 mmol) in DMF (10 mL) was added. The reaction was heated in an oil bath at 60 ° C for 2 hours and then cooled and poured into water and 5% potassium bisulfate. The resulting mixture was extracted with ethyl acetate (4 χ 250 mL). The combined organics were washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated. Flash column chromatography eluting with hexane / ethyl acetate 70/30 gave the title compound as a white foam, 13.9 g (33.4 mmol, 81% yield). LRMS (ESI) m / z 417 [(M + H); calcd for C22<sub>7</sub>H29N<sub>2</sub>Oh<sub>6</sub>: 417].
Step 6: (3R, 5S) -5- (Methoxycarbonyl) pyrrolidin-3-yl 4-vinyl-1,3-dihydro-2Hisoindole-2H-carboxylate hydrochloride
HCI
N CO, CH<sub>3 </sub>H
1- / tert-Butyl 2-methyl (2S ', 4R) -4 - {[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy] pyrrolidine-1,2-dicarboxylate solution 13.9 g, 33.4 mmol) in ethyl acetate (700 mL) was cooled in an ice bath and saturated with hydrogen chloride gas. The reaction vessel was closed genetically and allowed to warm to room temperature. After 3.5 h the solvent was removed by evaporation to give the title compound as a gray solid (11.2 g, 95% yield). 1 H NMR (500 MHz, ppm, CD 3)<sub>3</sub>OD) b 7.47-7.45 (m, 1H), 7.32-7.31 (m, 1H), 7.26-7.21 (m, 1H), 6.79-6.73 (m, 1H). 5.79 - 5.73 (m, 1H), 5.46 (s, 1H), 5.41 - 5.38 (m, 1H). 4.80 - 4.72 (m, 4H), 3.91 (s, 3H), 3.74 - 3.63 (m, 2H), 2.77 - 2.71 (m, 1H), 2.51-2.46 (m, 1H). LRMS (ESI) m / z 317 [(M + H)<sup>+</sup>; calc. for C17H21N2O4: 317],
Step 7: Methyl 2-dimethylpent-4-enyhoxy] carbonyl] -3-methyl-L-valyl- (4R) -4 {[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl oxy] -L-pronate
<img file="RS51101B_D0067.tif" />
51101Β
To a solution of (3R, 5Y) -5- (methoxycarbonyl) pyrrolidin-3-yl 4-vinyl-1,3-dihydro-2 H -indole-2-carboxylate hydrochloride (2.00 g, 5.67 mmol) and A- [ [(2,2-dimethylpent-4-enyl) oxy] carbonyl} -3-methyl-L-valine (1.54 g, 5.67 mmol) in DMF (100 mL) was added EDC (1.41 g, 7.37 mmol), HOBt (1.00 g) , 7.37 mmol) and DIPEA (3.16 mL, 22.8 mmol). The reaction mixture was stirred at room temperature for 18 hours and then diluted with ethyl acetate and aqueous NaHCO<sub>3</sub>. The layers were separated and the organic layer was washed with water and brine, dried over NaiSO 2. filtered and concentrated. The crude residue was purified on silica gel (eluting with a gradient of 5% to 50% ethyl acetate in hexanes) to give methyl A - {[(2,2-dimethylpent-4-enyl) oxy] carbonyl} -3-methyl-L-valyl. - ((4R) -4 - {[(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} -L-prolineate (2.75 g, 85% yield) as a white foam. LRMS (ESI) m / z 570 [(M + H) & lt; + & gt ;.<sup>+</sup>; calc. for C31H44N3O7: 570],
Step 8: Methyl (5R, 7S, 10S) -10-tert-butyl-15,15-dimethyl-3,9,12-trioxole, 6,7,9,10,11, 12,14,15,16- decahydro-5H-2,22: 5,8-dimethano-4,13,2,8,1,1-benzodioxatriazacycloicosine-7-carboxylate
<img file="RS51101B_D0068.tif" />
Methyl A - {[(2,2-dimethylpent-4-enyl) oxy] carbonyl} -3-methyl-L-valyl- (4R) -4 - {[4-vinyl-1,3-dihydro-2H-isoindol-2 solution] -yl) carbonyl] oxy} -L-proinate (2.46 g, 4.32 mmol) in anhydrous dichloromethane (450 mL) was purified with nitrogen for 15 min. A solution of bis (tricyclohexylphosphine) -3-phenyl-1H-indene-1-ylidenerutenium dichloride (Neolyst M1 catalyst obtained from Strem) (0.40 g, 0.43 mmol) in degassed. anhydrous dichloromethane (50 mL) was then added dropwise over 30 min. The reaction mixture was stirred at room temperature, during which 0.2 g of catalyst was added every approximately 8-12 hours. The progress of the reaction was monitored by HPLC until the reaction was complete at 48 hours. The residue was purified by flash chromatography on silica gel. eluting with 10-70% EtOAc / hexane to give methyl (5RJS, 10S) -10- / m<sup>,</sup>-butyl-15,15-dimethyl-3.9.12
51101 Β trioxo-1,6,7,9,10,11,1,1,1,14,15,16-decahydro-5H-2.22: 5,8-dimethano-4,13,2,8,1,1-benzodioxatriazacycloicosine-7-carboxylate (1.85 g, 76% yield). LRMS (ESI) mtz 542 [(M + H))<sup>+</sup>; calc. for C29H40N3O7: 542].
Step 9: (5R, 7S, 10SF10-tert-Butyl-15,15-dimethyl-3,9,12-trioxo-1,6,7,9,10,1,12,
14,15,16-decahydro-5H-2,22: 5,8-dimethano-4,13,2,8,1,1-benzodioxatriazacyclocycosine-7-carboxylic acid
<img file="RS51101B_D0069.tif" />
In a solution of methyl (5R, 7S, 10S) -10- / tert-butyl-15,15-dimethyl-3,9,12-trioxo-
1.6.7.9.10.11.12.14.15.16-decahydro-5H-2,22: 5,8-dimethano-1,1,1,2,8,11-benzodioxatriase cycloicosine-7-carboxylate (0.9 g, 1.67 mmol) in THF : H<sub>2</sub>O (2: 1, 45 mL) was added LiOH (0.40, 16.7 mmol). The reaction mixture was heated to 40 ° C and stirred for 1 hour. The reaction mixture was diluted with aqueous HCl and extracted with EtOAc. The combined EtOAc layer was washed with water, brine, dried over Na<sub>2</sub>SO4. filtered and concentrated. The product was used without further purification. LRMS (ESI) m / z 528 [(M + H)]<sup>4</sup>; calc. for C<sub>28</sub>l EgNjCB: 528],
Step 10: (5R, 7S, 10S) -10- (Grc-Butyl-N- (1R, 2S) -1 - {[(cyclopropylsulfonyl) amino | carbonyl} -2-vinylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo-1,6,7,9,10,11,11,14,15,16decahydro-5H-2,22: 5.8- dimethano-4,13,2,8,11-benzodioxatriazacycloicosin-7-carboxamide
Solution (5R, 7S, 10S) -10-tert-butyl-15,15-dimethyl-3,9,12-trioxo-1,6,7,9,10,11,12,14,
15.16-decahydro-5H-2.22: 5,8-dimethano-4,13,2,8,1-benzodioxatriazacycloicosine-7-carboxylic acid (100 mg, 0.19 mmol), (12? .25) -1 - {[(cyclopropylsulfonyl) ) amino | carbonyl} -2-imnylcyclopropanaminium chloride (Llinas-Brunet et al. US03 / 15755 and Wang et al. WO 03/099274) (76 mg, 0.28 mmol). O- (7-azabienzotriazol-1-yl) -N, N, N<sup>,</sup>.N'-tetramethyluiOnium
51101 Β Phosphorus hexafluoride (HATU, 108 mg, 0.28 mmol), DIPEA (0.073 mL, 0.42 mmol) and 4-dimethylaminopyridine (2 mg) in dichloromethane (5 mL) were stirred at 40 ° C for 1 hour. The reaction solution was diluted with aqueous saturated NaHCO 3 solution<sub>3</sub>, and extracted with EtOAc. The combined EtOAc layer was washed with water, brine, dried over Na<sub>2</sub>SO4, filtered and concentrated. The residue was purified by flash chromatography eluting with 3% MeON / SNgSE to give (5RJS.10S) -10-tert-butyl-A - ((1R, 2S) -1 {[(cyclopropylsulfonyl) amino] carbonyl) ] -2-vinylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo-
1,6,7,9,10,11,12,14.15,16-decahydro-5H-2.22: 5,8-dimethano-4,13,2,8,11-benzodioxatriase cycloicosine-7-carboxamide (80 mg, 57% yield). 1 H NMR (400 MHz, ppm, CDCl 3) δ 7.48 (s, 1H). 7.23 (s, 1H). 7.12 (d, 1H), 6.23 (d, J = 15.9 Hz, 1H), 5.94 (m, 1H), 5.76 (m, 1H), 5.50 (m, 2H), 5.43 (s, 1 H), 5.24 (d, J = 16.6 Hz, 1H). 5.11 (d, 1H), 4.70 (s, 2H), 4.61 (d, 1H), 4.48 (m, 3H), 4.35 (d, 1H), 4.14 (d, 1H), 3.74 d, 1H), 3.34 (d, 1H), 2.89 (m, 1H), 2.43 (dd, 2H). 2.06 (m, 1H), 1.93 m. 1 H). 1.89 (dd, 1H), 1.43 (d, 1H), 1.25 (m, 3H), 1.09 (s, 3H), 1.06 (s, 9H), 0.86 (s, 3H). LRMS (ESI) m / z 740 [(M + H)<sup>+</sup>; calc. for C<sub>37</sub>H<sub>5</sub>оН<sub>5</sub>09S: 740].
EXAMPLE 4 (5R, 7S, 10S) -10- / Grc-Butyl-A<sup>r</sup>- ((1R, 2S) -1- {i (cycloDronylsulfonyl) aminocarbonyl} -2vinylcyclopropyl) -3,9,12-trioxo-6,7,9,10,11,12,14J5,16,17-decahydro-1H , 5H-2,23.'5,8dimethano-4,13,2,8,11-benzodroxatriazacyclohenicosine-7-carboxamide (ΙΙΙ-12)
<img file="RS51101B_D0070.tif" />
The title compound was prepared according to the procedure used for EXAMPLE 3 except that 3-methyl-A - [(hex-5-enyloxy) carbonyl] -L-valine (prepared according to the procedure below) was used. instead of N - {[(2,2-dimethylpent-4-phenyl) oxy] carbonyl] -3-methyl-L-valine in Step 7. 1 N NMR (500 MHz, ppm. CD3OD) b 9.13 (s, 1H), 7.26 (t, 1H), 7.23 (d, 1H), 7.16 (d, 1H), 6.39 (d. 16.4 Hz, 1H). 6.08 (m, 111),
5.76 (m, 1H), 5.38 (s, 1H), 5.29 (d, 1H), 5.12 (d, 1H). 4.79 (d, 1H), 4.73-4.63 (m, 4H), 4.41 (s, 1H). 4.37 (q, 1H), 4.24 (d, 1H). 3.96 (dd, 1H), 3.77 (quin., 1H). 2.94 (m, 1H). 2.51
51101Β (q, 1 Β), 2.29 - 2.13 (m, 4 Β). 1.87 (dd. 1 Η), 1.68 (m, 2 (), 1.53 (quin., 2 Η), 1.44 (dd, 1 Η). 1.25 (m, 2 (), 1.05 (s, 9 (). LRMS (ESI) m / z 726 [(M + H)<sup>+</sup>; calc. for C<sub>36</sub>H<sub>48</sub>N<sub>s</sub>Oh<sub>9</sub>S: 726],
EXAMPLE 5 (5R, 7SJ0SM0-Butyl-A - ((1R, 2S) -1- {i (cisopropylsulfonyl) aminoylcarbonyl) -2vinylcyclopropyl) -3,9,12-trioxo-6,7,9,10,11, 12,14,15,16,17-decahydro-1H, 5H-2,23: 5,8dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosine-7-carboxamide (W-133)
<img file="RS51101B_D0071.tif" />
The title compound prepared according to the procedure used for EXAMPLE 3 with the difference that 3-methyl-A - [(hex-5-emyloxy) carbonyl] -L-norleucine (prepared according to the procedure below) was used instead N - {[(2,2-dimethylpent-4-phenyl) oxy] carbonyl) -3-methyl-L-valine in Step 7. 1 N NMR (500 MHz, ppm. CD<sub>3</sub>OD) b 7.24 (t, 1H), 7.23 (d, 1H), 7.15 (d, 1H), 6.91 (d, 1H), 6.37 (d, J = 16.1 Hz, 1H), 6.07 m, 111), 5.75 (m, 1H), 5.39 (s, 1H), 5.29 (d, 1H), 5.12 (d, 1H), 4.77 (d, 1H), 4.66 (m, 3 H), 4.57 (m, 1H), 4.47 (q, 1H), 4.39 (q, 1H), 4.27 (d, 1H), 3.90 (dd, 1H), 3.77 (quin., 1H) ), 2.96 (m, 1H), 2.49 (q, 1H), 2.29 (m, 1H), 2.22 (m, 3H), 1.88 (dd, 1H), 1.75 (m, 2H). 1.64 (m, 2H), 1.52 (m, 2H), 1.39 (m, 5H), 1.27 (m, 1H), 1.18 (m, 1H). 1.09 (m, 2H), 0.94 (t, 3H). LRMS (ESI) m / z 726 [(M + H)<sup>+</sup>; calc. for C36H<sub>48</sub>N<sub>5</sub>Oh<sub>9</sub>S: 726],
51101 Β
EXAMPLE 6 (5R, 7S40S) -10-Butyl-WRR, 2S) -1- {Cyclopropylsulfonyl) aminocarbonyl-2-vinyl] cyclopropyl) -3,9,12-trioxo-1,6,7,9,10,11,12 , 14,15,16,17,18-dodecahydro-5H-2,24: 5,8dimethano-4,13,2,8,11-benzodioxatriazacycloiodocosine-7-carboxyanide (III-I98)
<img file="RS51101B_D0072.tif" />
W-198
The title compound was prepared according to the procedure used for EXAMPLE 3 except that № [(hept-6-en-1-yloxy) carbonyl] -L-norleucine (prepared according to the procedure below) was used instead of N - {[(2,2-dimethylpent-4-phenyl) oxy] carbonyl} -3-methyl-L-valine in Step 7. 1 N NMR (500 MHz, ppm. CD<sub>3</sub>OD) b 9.26 (s, 1H), 7.39 (d, 1H), 7.24 (t, 1H). 7.15 (d, 1H), 6.30 (d, J = 15.9 Hz, 1H), 6.20 (m, III),
5.75 (m, 1H), 5.53 (s, 1H), 5.31 (d, 1H), 5.12 (d, 1H), 4.70 (m, 4H), 4.43 (dd, 1H). 4.34 (m, 2H), 4.27 (q, 1H), 3.91 (dd, 1H), 3.79 (quin., 1H). 3.31 (m, 1H), 2.97 (m, 1H), 2.31 (m, 1H), 2.22 (m, 3H), 1.89 (dd, 1H), 1.74 (m, 2H). 1.66 (m, 1H), 1.56 (m, 3H). 1.38 (m.
H). 1.19 (m, 1H), 1.09 (m, 2H). 0.94 (p. 3 II). LRMS (ESI) m / ~ 740 [(M + H) & lt; + & gt;]<sup>+</sup>: calc. for C37H50N5O9S: 740],
51101Β
EXAMPLE 7 (5R, 7S, 10S) -10-tert-Butyl-N - ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl] -2vinylcyclopropyl) -15,15-dimethyl-3,9, 12-thioxo-6,7,9,10,11,12,14,15,16,17-decahydrolH, 5H-2,23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosin- 7-carboxamide (W-199)
<img file="RS51101B_D0073.tif" />
W-199
The title compound was prepared according to the procedure used for EXAMPLE 3 except that A - {[(2,2-dimethylhex-5-enyl) oxy] carbonyl} -3-methyl-L-valine was prepared according to the method used. is given below) instead of: V - {[(2,2-dimethylpent-4-phenyl) oxy] carbonyl} -3-methyl-L-valine in Step 7. <sup>1</sup>11 NMR (500 MHz, ppm, CD 3 OD) δ 9.17 (s, 1H), 7.27 (t ../= 7.5 Ηζ. 1 H), 7.21 (t, ./ = 7.5 Hz, 2 H). 7.16 (d, J = 7.5 Ηζ, 1 H), 6.38 (d, ./= 16 Hz, 1 H), 6.03 (m, 1 H), 5.79 (m, 1 H), 5.32 (m, 2 II) , 5.13 (m, 1H), 4.82-4.77 (m, 1H), 4.73-4.61 (m, 4H), 4.48 (s, 1H), 4.39 (m, 1H). 4.19 (d. .7 = 12 Hz. 1H), 3.96 (m, 1H), 2.96 (m, 1H), 2.59-2.55 (m, 1H), 2.35-2.12 (m, 4H), 1.89 (m, 1H), 1.49-1.23 (m, 6H), 1.51-0.98 (m, 14H), 0.95-0.85 (m, 4H). LRMS (ESI) m / z 754 [(M + H) & lt; + & gt ;; calc. for C3SH52N5O9S: 754].
51101 Β
EXAMPLE 8 (5R, 75,10S) -10-tert-Butyl N - ((1R, 2R) -1- {1 (cyclopropylsulfonyl) amino] carbonyl) -2-ethylcyclopropyl) -3,9,12-trioxo-1, 6,7,9,10,11,12,14,15,16,17,18-dodecahydro-5H-2,22: 5,8dimethano-4,13,2,8,11-benzodioxatriazacyclocycosine-7-carboxamide ( ΙΠ-200)
<img file="RS51101B_D0074.tif" />
A solution of EXAMPLE 2 (0.32 mg, 0.45 mmol) and palladium on carbon (10% w / w, 0.03 g) in EtOAc (10 mL) was stirred vigorously under a hydrogen balloon for 1 hour. The reaction mixture was filtered and concentrated. The residue was purified by reverse phase HPLC (DeltaPak C18 column), with 40-65% CH<sub>3</sub>CN in water (with NH<sub>4</sub>OAc 1 g / L). Fractions were concentrated, diluted with aqueous saturated IaNSO 2 solution (20 mL) and extracted with CH<sub>2</sub>C1<sub>2</sub> (3 x 70 mL). Combined CH<sub>2</sub>C1<sub>2</sub> the layers were washed with water (50 mL), dried over Na<sub>2</sub>SO<sub>4</sub>, filtered and concentrated to give (5R, 7S, 10S) -10-to'c-butyl-N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) aminocarbonyl} -2-ethylcyclopropyl) -3, 9.12-Trioxo-1,6,7,9,10,11,12.14,15.16.17,18 dodecahydro-5H-2,22: 5,8-dimethano-4,1,2,2,8,1 l-benzodioxatriazacycloicozin-7-carboxamide ( 0.31 g, 97% yield). 1 H NMR (CD<sub>3</sub>OD ppm) b 7.23 (t, 1H). 7.14 (d, 1H), 7.10 (d, 1H). 7.02 (d, 1H), 5.52 (s, 1H), 4.74 - 4.60 (m, 4H), 4.48 - 4.30 (m, 4H), 3.88 (d, 1H), 3.75 (s, 1H), 2.99 (m, 1H). 2.62 (m, 1H), 2.41 (m, 2H), 2.14 (m, 1H), 1.79 (m, 1H), 1.65 - 1.51 (w, 6H), 1.47-1.19 (m, 5H) , 1.07 (s, 9 (), 0.99 (t, 3H). LRMS (ESI) m / z 716 [(M + H)<sup>+</sup>; calc. for C35H<sub>5</sub>оН<sub>5</sub>0<sub>9</sub>S: 716).
51101 Β
EXAMPLE 9 (5R, 7S, 10S) -10-tert-Butyl-N - ((1R, 2R) -1- {Rcyclopropylsulfonyl) amino | carbonylF2ethylcyclopropyl) -3,9,12-frioxo-6,7,9<sub>t</sub>10,11,12,14,15,16,17,18,19-dodecahydro-1H, 5H2,23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosine-7-carboxamide (ΙΙΙ- 201)
<img file="RS51101B_D0075.tif" />
W-201
The title compound was prepared from EXAMPLE 4 using the procedure described for EXAMPLE 8. 1 N NMR (500 MHz, ppm, CD<sub>3</sub>OD) b 7.23 (t, 1H), 7.14 (d, 1H), 7.10 (d, 1H), 7.02 (d, 1H), 5.36 (s, 1H), 4.71 (m, 3H) , 4.64 (t, 1H). 4.56 (m, 1H). 4.40 (m, 2H), 4.24 (d, 1H), 3.96 (dd, 1H), 3.72 (quin. 1H), 2.98 (m, 1H). 2.58 (m, 1H), 2.49 (m, 2H),
2.15 (t, 1H), 1.69 -1.19 (m, 15H), 1.09 (m, 1H), 1.06 (s, 9H), 0.98 (t, 3H). LRMS (ESI) m / z 730 [(M + H)<sup>+</sup>; calc. for [^ lfeNAS: 730].
EXAMPLE 10 (SRJSJOSElO-Butyl-A-R1R ^ R1-1-cyclopropylsulfonylaminocarbonine-letylcyclopropyl) -3,9,12-trioxo-6,7,9,10,11,1,1,14,15,16,17,18,19 -dodecahydro-1H, 5H ~ 2.23: 5,8-dimethano-4,13,2,8<sub><</sub>11-Benzodioxatriazacyclohenicosine-7-carboxamide (W-202)
<img file="RS51101B_D0076.tif" />
W-202
The title compound was prepared from EXAMPLE 5 using the procedure described for EXAMPLE 8. 1 N NMR (500 MHz, ppm, CD<sub>3</sub>OD b 7.23 (t 1H). 7.14 (d, 1H), 7.09 (d, 1H). 6.99 (d, 1H), 5.39 (s, 1H), 4.76 - 4.61 (m, 4H). 4.43 (m, 3 II). 4.29 (d, 1H), 3.92
51101 Dd (dd, 1 Η), 3.69 (quin., 1 Β), 2.99 (m. 1 Η), 2.57 (m, 1 Η), 2.51 (m, 2 Η), 2.19 (tt, 1 Η), 1.77 (m, 1 Η), 1.70 -1.30 (m, 20 Η), 1.17 (m, 2), 1.10 (m, 2), 0.99 (t, 3), 0.95 (t, 3 Η). LRMS (ESI) m / z 730 [(M + H)<sup>+</sup>; calc. for C<sub>36</sub>H<sub>5</sub>zN<sub>5</sub>Oh<sub>9</sub>S: 730],
EXAMPLE 11 (5R, 7S, 10S) -10-Butyl- N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino] carbonyl-2-ethylcyclopropyl) -3,9,12-trioxo-1,6, 7,9,10,11,1,1,14,15,16,17,18,19,20-tetradecyclohydro 5 H2,24: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclodocosin-7 -carboxamide (W-203)
<img file="RS51101B_D0077.tif" />
The title compound was prepared from EXAMPLE 6 using the procedure described for EXAMPLE 8. 1 N NMR (500 MHz, ppm. CD<sub>3</sub>OD) b 7.2 (m, 1 H), 7.15 (d, 1 H), 7.11 (d, 1 H), 5.55 (s, 1 H), 4.70 (m, 4 H). 4.49 (m, 1H), 4.38 (t, 1H), 4.29 (m, 2H), 3.94 (dd, 1H), 3.73 (quin., 1H), 3.00 (m, 1H), 2.63 (quin, 1H), 2.51 (m, 1H), 2.38 (m, 1H), 2.20 (tl, 1H), 1.76 (quin., 1H), 1.68-1.07 (m, 24H). 1.00 (t, 3H), 0.95 (t, 3H). LRMS (ESI) m / z 744 [(M + H))<sup>+</sup>; calc. for C<sub>37</sub>H<sub>5</sub>4N<sub>5</sub>Oh<sub>9</sub>S: 744],
51101 Β
EXAMPLE 12 (5R, 7S, 10S) -10-tert-Butyl] -N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino] carbonyl) -2-ethylcyclopropyl) -15,15-dimethyl-3,9 , 12-trioxo-1,1,7,7,9,10,11,1,1,14,15,16,17,18-dodecahydro5H-2,22: 5,8-dimethano-4,13,2,8,11 -benzodioxatriazacycloicosin-7-carboxamide (W-204)
<img file="RS51101B_D0078.tif" />
The title compound was prepared from EXAMPLE 3 using the procedure described for EXAMPLE 8. <sup>[</sup>1 H NMR (400 MHz, ppm, CD<sub>3</sub>OD) b 9.06 (s, 1H), 7.22 (dd, 1H), 7.13 (d. 1H), 7.07 (d, 1H), 5.51 (s, 1H). 4.72 (d, 2H), 4.68 (d, 2H), 4.44 (d, 2H), 4.28 (m, 2H). 3.87 (dd, 1H), 3.28 (m, 1H), 2.98 (d, 1H), 2.85 (m, 3H). 2.52 (m, 1H), 2.43 (m, 2H). 2.15 (m, 1H), 1.15-1.17 (m, 3H), 1.41 (m, 2H). 1.30 (m, 1H), 1.21 (m, 4H), 1.08 (m, 1H), 1.06 (s, 3H), 1.05 (s, 9H), 0.98 (t, 3H), 0.81 pp. 3 H). LRMS (ESI) m-744 [(M + H); calc. for C37H54N5O9S: 744].
EXAMPLE 13 (5R, 7S, 10S) -10- (Grc-Butyl-N - ((1R, 2R) -1 - {| (cyclopropylsulfonyl) aminocarbonyl-2-ethylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo -6,7,9,10,11,11,14,15,16,17,18,19dodecahydro-1H, 5H-2,23: 5,8-dimethano-4,13,2,8,11 - benzodioxatriazacyclohenicosin-7carboxamide (W-205)
<img file="RS51101B_D0079.tif" />
W-205
51101 Β
The title compound was prepared from EXAMPLE 7 using the procedure described for EXAMPLE 8. 1 N NMR (500 MHz, ppm, CD<sub>3</sub>OD b 9.09 (s, 1H), 7.24 (t, J = 7.5 Hz. 1H), 7.15 (d. ./ = 7.5 Hz. 1 II), 7.10 (d, J - 7.5 IIz, 1 II). 5.53 (s, 1 II), 4.75 - 4.59 (m, 4 11), 4.44 - 4.37 (m, 3 H). 4.20 (d, J = 12 Hz, 1H). 3.95 - 3.91 (m, 1H), 3.31 (m, 2H), 2.99 - 2.96 (m, 1H), 2.62 - 2.46 (m, 3H), 2.17 - 2.13 (m, 1H), 1.67 - 1.50 (m, 6H), 1.37-1.18 (m, 7H),
1.15 - 0.96 (m, 16H), 0.80 (s, 3H). LR.MS (ESI) m / z 758 [(M + H))<sup>+</sup>; calc. for SLJVD: 758],
Alternative preparation:
Step 1: 1-Bromo-2,3-bis (bromomethyl) benzene
Vg Br
To a suspension of 3-bromo-o-xylene (999 g, 5.40 mol) in chlorobenzene (9 L) at room temperature was added A-bromosuccinimide (1620 g, 9.1 mol) and benzoyl peroxide (2.6 g, 10.8 mmol). The reaction mixture was heated to 80 ° C and stirred under nitrogen for 18 hours. The reaction mixture was cooled to 70 ° C and another portion of NBS (302 g, 1.7 mol) was added. The reaction mixture was heated to 80 ° C and stirred under nitrogen for 22 hours. The reaction mixture was cooled to room temperature, diluted with heptane (6 L) and filtered, the filter cake was washed with heptane (4 L) and the combined filtrates were evaporated. The crude product was dissolved in heptane (2 L) and chloroform (200 mL) and filtered through basic aluminum trioxide (500 g). The aluminum trioxide plug was washed with heptane (4 L) and the combined liltrates were evaporated to give 1-bromo-2,3bis (bromomethyl) benzene (1760 g, crude weight) which was used without further purification. 1 H NMR (CDCl 3) δ (ppm) 7.56 (d, J = 8.0 Hz, 1H), 7.31 (d, 8.0 Hz, 1H), 7.26 (s, 1H),
7.16 (t, J = 8.0 Hz. 1H), 4.84 (s, 2H), 4.64 (s, 2H).
51101 Β
Step 2: 2-Benzyl-4-bromoisoindoine hydrochloride
<img file="RS51101B_D0080.tif" />
Potassium bicarbonate (657 g, 6.56 mol) was suspended in MeCN (17 L) and the mixture was heated to 80 ° C. Solutions of crude 1-bromo-2,3-bis (bromomethyl) benzene (900 g, 2.63 mol in 1 L MeCN) and benzylamine (281 g, 2.63 mol in 1 L MeCN) were added at the same time. . The reaction mixture was stirred at 77 ° C for 2 hours and then cooled to room temperature and stirred for 16 hours. The contents of the reaction vessel were cooled, filtered and the solvent was removed by evaporation. The reaction was partitioned between water (6 L) and EtOAc (2 L). The pH was adjusted to pH> 9 by adding 1M K<sub>2</sub>In SO 2, the layers were separated and the aqueous phase was extracted with additional EtOAc (2 L). The combined organics were washed with brine. dried with anhydrous Na<sub>2</sub>SO<sub>4</sub>, filtered and evaporated. The crude oil was diluted with EtOH (300 mL) and cooled to 0 ° C. Methanolic HCl was added until the mixture became acidic, then MTBE (700 mL) was added and the mixture was sonicated, then stirred for 15 hours. MTBE (1 L) was added and the mixture was filtered and washed with 20% EtOH in MTBE and then with MTBE. The solid was air dried to give 2-benzyl-4-bromoisoindoline hydrochloride (211 g). An additional portion of the product (86 g) was isolated by concentrating the mother liquors. LRMS (ESI) m / z 289 [(M + H) & lt; + & gt ;; calc. for C<sub>15</sub>H<sub>15</sub>BrN: 289],
Step 3: 4-Bromoisoindoline
<img file="RS51101B_D0081.tif" />
To a solution of 2-benzyl-4-bromoisoindoline hydrochloride (11 g, 30.96 mmol) in 200 mL of EtOAc was added 1M NaOH (100 mL) and the mixture was stirred for 30 min. The organic layer was separated, washed with brine. dried over anhydrous Na<sub>2</sub>SO<sub>4</sub> and the solvent was evaporated to
51101Β of oil which was azeotroped once with toluene (50 mL). The oil was dissolved in chlorobenzene (50 mL) and 4A molecular lattice (5 g) was added to the stirred solution. After 10 min, 1-chloroethyl chloroformate (5.6 mL, 51 mmol) was added dropwise over 5 min. The reaction mixture was then heated to 90 ° C for 2 hours, cooled to room temperature and filtered. The solids were washed with chlorobenzene (5 mL) and methanol (40 mL). The filtrate was heated to 70 ° C for 1 hour, allowed to cool and stirred at room temperature overnight. The solids were filtered, washed with chlorobenzene (2 mL) and hexane and dried to give 6.84 g of the title compound. LRMS (ESI) m / z 198.1 [(M + H) & lt; + & gt ;.<sup>+</sup>; calc. for C<sub>8</sub>H<sub>9</sub>BrN: 198.0],
Step 4: 1-t-Butyl 2-methyl (2S, 4R) -4-H (4-bromo-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy] pyrrolidine-1,2-dicarboxylate
To a solution of (2S, 4R) -BOC-4-hydroxyproline methyl ester (126.3 g, 515 mmol) in DMF (960 mL) at 0 ° C was added N, N'-carbonyldiimidazole (83.51 g, 515 mmol). The reaction mixture was stirred at room temperature for 3 hours. 4-Bromoisoindoline hydrochloride (120 g, 515 mmol) and diisopropylethylamine (96.3 mL, 540 mmol) were added and the reaction mixture was warmed to 50 ° C for 6 hours and then allowed to cool to room temperature and stirred overnight. The reaction mixture was partitioned between EtOAc (3 L) and 10% aqueous KHSO<sub>4</sub> (6 L), the aqueous layer was re-extracted with EtOAs (2 L) and the combined organic phases were washed with 10% aqueous NaHCO 3.<sub>3</sub>, saline, dried over Na<sub>3</sub>SO<sub>4</sub> and the solvent was evaporated to a foam (239 g). LRMS (ESI) m / z 471.0 [(M + H) & lt; + & gt;]<sup>+</sup>; calc. for C ^ oFEfiBrlSBOf ,: 471.1].
51101Β
Step 5: 1-t-Butyl 2-methyl (2S, 4R) -4- {N4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy] pyrrolidine-1,2-dicarboxylate
<img file="RS51101B_D0082.tif" />
To a solution of 1-t-butyl 2-methyl (2S, 4R) -4 - {[(4-bromo-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy} pyrrolidine-1,2-dicarboxylate g, 21.3 mmol) in ethanol (200 mL) was added potassium vinyl trifluoroborate (4.28 g, 32 mmol) and Iriethylamine (4.5 mL, 32 mmol) followed by dichloro [1,1-bis (diphenylphosphino) ferrocene] palladium (II) chloride dichloromethane adduct (175 mg, 0.21 mmol). The reaction mixture was heated to reflux for 6 hours, cooled to room temperature, diluted with 10% aqueous KHSO.<sub>4</sub> and ethanol was removed by evaporation in vacuo. The aqueous residue was extracted with EtOAc and the organic phase was washed with brine, dried over Na<sub>2</sub>SO<sub>4</sub>, the solvent was evaporated and the crude product was purified by chromatography on silica eluting with 40-60% EtOAc / hexane to give after evaporation the title compound (8.18 g), LRMS (ESI) m / z 417.2 [(M + H)]. '; calc. for C<sub>22</sub>1I<sub>29</sub>N<sub>2</sub>Oh<sub>6</sub>: 417.2].
Step 6: (3R, 5R) -5- (Methoxycarbonyl) pyrrolidine-3-14-vinyl-13-dihydro-2H-isoindole-2-carboxylate hydrochloride
<img file="RS51101B_D0083.tif" />
Mixture of 1-t-butyl 2-methyl (2S.4R) -4 - [([(4-vinyl-1,3-dihydro-2H-isoindol-2-yl) carbonyl] oxy) pyrrolidine-1,2-dicarboxylate 43.2 mmol) and HCl / dioxane (4 M) (43.2
51101Β mL, 173 mmol) was stirred at room temperature for 2 hours. The reaction mixture was concentrated to remove dioxane, followed by concentration from Et<sub>2</sub>O to give (3R, 5S) -5 (methoxycarbonyl) pyrrolidin-3-yl 4-vinyl-1,3-dihydro-2H-isoindole-2-carboxylate hydrochloride as an off-white solid (15 g) which was used without further purification. LRMS (ESI) m / z 317 [(M + H)]<sup>+</sup>: calc. for Ci<sub>7</sub>H<sub>21</sub>N<sub>2</sub>Oh<sub>4</sub>: 317J.
Step 7: Methyl N] [(2,2-dimethylhex-5-en-1-yl) oxy] carbonyl} -3-methyl-L-valyl- (47?) 4 - {[(4-vinyl-1, 3-dihydro-2Z7-isoindol-2-yl) carbonyl] oxy] -L-prolineate
<img file="RS51101B_D0084.tif" />
To a solution of (3R, 5S) -5- (methoxycarbonyl) pyrrolidin-3-yl 4-vinyl-1,3-dihydro-2H-isoindole-2-carboxylate hydrochloride (5.0 g, 14.2 mmol) and V - {[(2 , 2-dimethylhex-5-enyl) oxy] carbonyl} -3methyl-L-valine (4.0 g, 14.2 mmol) in DMF (20 mL) at room temperature was added DIPEA (2.5 mL, 14.2 mmol), EDC (5.5 g , 28.4 mmol) and HOAt (1.9 g, 14.2 mmol). After 18 h, the reaction mixture was poured into Et<sub>2</sub>O, and extracted with 1 N HCl The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with 1 N HCl, water, IaNSO 2 and brine. The organic layer was dried over MgSO 4<sub>4</sub> and the solvent was removed in vacuo. The crude product was purified on silica (30% EtOAc in hexanes) to give 4.2 g of the title compound as a thick oil. LRMS (ESI) m / z 584.4 [(M + H) & lt; + & gt;]<sup>+</sup>; calc. for C32H46N3O7: 584.3],
51101Β
Step 8: Methyl (57?, 75,105.18 lbs<sup>T</sup>) -10- [tert-butyl-15,15-dimethyl-3,9,12-trioxo 6,7,9,10,11,1,1,14,15,16,17-decahydro-m, 5,2,23 : 5,8-dimethano-4,13,2,8,1,1-benzodioxafriazacyclohenicosine-7-carboxylate
<img file="RS51101B_D0085.tif" />
To a solution of methyl V - [[(2,2-dimethylhex-5-en-1-yl) oxy] carbonyl} -3-methyl-L-valyl- (4R) -4 {[(4-vinyl, 3 -dihydro-2H-isoindol-2-yl) carbonyl] oxy} -L-prolineate (4.7 g, 8.05 mmol) in degassed (nitrogen bubble burst for 30 min) DCM (1410 mL) was added Zhan 1B catalyst (Zhan Catalyst 1B, RC-303, Zannan Pharma Ltd.) (0.591 g, 0.805 mmol). The mixture was then stirred at room temperature under an atmosphere of N<sub>2</sub>. After 7 p.m. the reaction was complete and DMSO (57 pL, 0.805 mmol) was added. The mixture was stirred for 2 hours and the mixture was concentrated in vacuo to -70 mL. The crude product was then directly purified on silica (gradient elution, 0-50% EtOAc in hexanes) to give 4.4 g of the title compound as an oil. LRMS (ESI) m / z 556.3 [(M + NI: calcd.
Step 9: Methyl (5 / ?, 7<sub>t</sub>S ', 100<sup>r</sup>1H-tert-butyl N, 5,15-dimethyl-3,9,12-trioxo-6,9,9,10,11,1,1,14,15,16,17,18,19-dodecahydro-1 H, 5fl -2,23: 5,8-d »netano-4,13,2,8,1,1-benzodioxatriazacycloquinosine-7-carboxylate
<img file="RS51101B_D0086.tif" />
In a solution of methyl (5 £, 7S, 10LS ', 118 £) -10- [t] butyl-1,15,15-dimethyl-3,9,12-trioxo6,7,9,10,11,12.14. 15,16,17-decahydro-1 / 7.5 / 7-2.23: 5,8-dimethano-4,13.2,8,11benzodioxatriazacyclohenicosine-7-carboxylate (4.4 g, 7.92 mmol) in EtOAc (79 mL) was added
51101 Β
Pd / C (0.421 g, 0.396 mmol). The Nj balloon was then placed on the reaction vessel. The vessel was quickly emptied and filled with Hk After 17 hours, the reaction was complete as determined by LC-MS. Pd / C was filtered through glass wool, and the crude product was purified on silica (elution gradient, 0-60% EtOAc in hexanes) to give 4.01 g of the title compound as a white powder. LRMS (ESI) m / z 558.4 [(M + H) & lt; + & gt;]<sup>+</sup>; calc. for C30H44N3O7: 558.3],
Step 10: (£ 5, 75<sup>,</sup>, 105) -10-fer<sup>,</sup>-ButyR5,15-dimethyl-3,9,12-trioxo-6,7,9,10,11,11,14,15,16,17,18,19-dodecahydro-1fl<sup>r</sup>, 5H-2.23: 5,8-dimethano-4,13,2,8,1,1-benzodioxatriazacyclohenicosine-7-carboxylic acid
<img file="RS51101B_D0087.tif" />
n G <sup>ο</sup>γ<sup>Ν</sup>go <sup>0</sup>
To a solution of methyl (5R, 7X10,9) -10-tov-butyl-15,15-dimethyl-3,9,12-trioxo-6,7,9.10,11,12, 14,15.16.17,18.19-dodecahydro -S.5 // - 2.23: 5,8-dimethano-4,13.2.8.11-benzodioxatriase cyclohenicosine-7-carboxylate (5.76 g, 10.33 mrnol) in THF (41.3 mL), MeOH (41.3 mL) and water (20.7 mL) at room temperature was added LiOH (4.33 g, 103 mmol). After complete conversion (45 min), as determined by LC-MS, the reaction was worked up by partitioning between Et<sub>2</sub>O and 1N HCl. The aqueous layer was then extracted with EtOAc. The combined organic layers were dried over MgSO 4<sub>4</sub> and the solvent was removed in vacuo to give 5.53 g of the title compound, which was used without further purification. LRMS (ESI) m / z 544.4 [(M + H) & lt; + & gt ;.<sup>+</sup>: calc. for S<sub>29</sub>N<sub>42</sub>^ O<sub>7</sub>: 544.3],
51101Β
Step 11: (5R, 7S, 10S) -10- (Grc-Butyl-N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino | carbonyl} -2-ethylcyclopropyl) -15,15-dimethyl- 3,9,12-trioxo-6,7,9,10,11, 12.14,15,16,17,18,19dodecahydro-1H, 5H-2,23: 5,8-dimethano-4,13,2, 8,11-benzodioxatriazacyclohenicosin-7carboxamide (III-205)
To a solution of (5R, 7S, 105) -10- [t-Butyl-1,15-dimethyl-3,9,12-trioxo-6,7,9,10,11,12,
14,15J6,17,18<sub>}</sub>19-Dodecahydro-1 / 7.57 / -2.23: 5,8-dimethano-4,13,2,8.11-benzodioxatriase cyclohenicosine-7-carboxylic acid (5.53 g, 10.17 mmol) and (17?, 2J) - DIPEA (7.11 mL, 40.7 mmol) and IIATU (5.03 g, 13.22 mmol) were added to 1-amino-, V (cyclopropylsulfonyl) -2-ethylcyclopropanecarboxamide hydrochloride (3.28 g, 12.21 mmol) in DMF (50.9 mL). After complete conversion (1 h), the reaction mixture was partitioned between EtOAc and 1N HCl. The organic layer was washed with brine three times, dried over MgSO 4<sub>4</sub>, and the solvent was removed in vacuo. The crude material was then purified on silica (gradient elution, 2080% EtOAc in hexanes) to give 5.8 g of the title compound as a white powder.
EXAMPLE 14 (5R, 7S, 10S) -10-tert-Butyl-A - ((1R, 2S) -1-f (cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropyl) -3,9,12-trioxo-1,6 7,9,10,11,1,1,1,14,15,16,17,18-dodecahydro-5H-2,22: 5,8dimethano-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxamide (N- 5)
<img file="RS51101B_D0088.tif" />
51101Β
Step 1: Methyl (5R, 7S, 10S) -10-tert-butyl-3,9,12-trioxo-1,6,7,9,10,11,12,14,15,16decahydro-5H-2,22: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacycloicosine-7-carboxylate
<img file="RS51101B_D0089.tif" />
Methyl (5R, 7S, 10S) -10- [t] butyl-3,9,12-trioxo-1,6,7,9,10.11,12,14.15,16-decahydro5H-2,22: 5, 8-Dimethano-4,1,2,2,1,1-benzodioxatriazacycloicosine-7-carboxylate was prepared according to the procedure used for methyl (5R, 7S, 10S) -10- [e] -c'-butyl-15 1,15-dimethyl-3,9,12-trioxo-1,6,7,9,10,11,1,1,14,15,16-decahydro-5H-2,22: 5,8-dimethano-4,13,2 , 8,11b-benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 3, Step 8) with the difference that 3-ethyl-N- (pent-4-enyloxy) carbonyl N-valine (prepared according to the procedure given below) was used instead of A- {f (2,2-dimethylpent-4- enyl) oxy] carbonyl} -3-methyl-L-valine in Step
7. LRMS (ESI) m / z 514 [(M + H)]<sup>+</sup>; calc. for C<sub>27</sub>H<sub>36</sub>N<sub>3</sub>Oh<sub>7</sub>: 514].
Step 2: Methyl i5R, 75,10S) -10-butyl-3,9,12-trioxo-1,6,7,9,10,11,1,1,14,15, 16,17,18-dodecahydro- 5H-2.22, 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclocycosine-7-carboxylate
<img file="RS51101B_D0090.tif" />
In a solution of methyl (5R, 7S.10S) -10- [m'-butyl-3.9.12-trioxo-1.6,7.9.10.11,12. 14,15,16,17,18-dodecahydro-5H-2.22: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacycloikosin-
7-Carboxylate (0.10 g, 0.20 mmol) in ethyl acetate (7 mL) was added 10% palladium on carbon (0.01 g). The reaction mixture was stirred under a hydrogen balloon for 5 hours at room temperature. The contents of the reaction vessel were filtered through celite and the filtrate was evaporated. Raw
51101 Β the product was used without further purification (0.09g, 90% yield). LRMS (ESI) m / z 516 [(M + H)<sup>+</sup>; calc. for C27H38N3O7: 516],
Step 3: ((5R, 7SJ0S) -10-tert-Butyl-N - ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl}} - 2-vinylcyclopropyl) -3,9,12-trioxo- 1,1,7,7,9,10, H, 12,14,15,16,17,18-dodecahydro5H-2,22: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacycloicosin-7- carboxamide
To a solution of methyl (5R, 7S, 10S) -10-to'c-butyl-3,9,12-trioxo-1,6,7.9.10, l 1.12, 14,15,16,17.18-dodecahydro-5H- 2.22: 5,8-dimethano-4,13,2.8,11-benzodioxatriazacycloicosine 7-carboxylate (90 mg, 0.18 mmol) in THF (2 mL) and MeOH (0.5 mL), LiOH (1N 1.75 mL) was added , 1.75 mmol). The reaction mixture was heated to 40 ° C and stirred for 1 hour, when complete consumption of methyl ester starting material was observed by LCMS. The mixture was then treated with 0.5 N HCl and EtOAc. The organic layer was then dried over K2CO3, and the solvent was removed in vacuo. The crude product was poured into DMF (1 mL).
To the above solution was added (1R, 2S) -1 - [[(cyclopropylsulfonyl) aminocarbonyl] -2-vinylcyclopropanaminium chloride (51 mg, 0.19 mmol), TBTU (77 mg, 0.24 mmol) and DIPEA (0.07 mL, 0.40 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was directly purified by reverse phase HPLC to give (5R, 7S, 10S) * 10- / erc * butyl-A - ((1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} - 2vinylcyclopropyl) -3,9,12-trioxo-1,6,7,9,10,11,1,1,14,15,16,17,18-dodecahydro-5H-2,22: 5,8dimethano-4,13 , 2,8,1 l-benzodioxatriazacycloicosine-7-carboxamide (34 mg, 28% yield). 11 NMR (500 MHz, ppm, CD 3 OD) δ 9.14 (s, 1H), 7.23 (t, 1H). 7.13 (d, 1H). 7.10 (d, 1H), 5.75 (quin., 1H), 5.53 (s, 1H), 5.29 (d, 1H), 5.12 (d, 1H), 4.75 - 4.59 (m, 5H), 4.42 (m, 2H). 4.34 (s, 1H), 4.30 (d, 1H), 3.88 (dd, 1H), 3.75 (m, 1H), 3.60 (q, 2H), 2.95 (m, 1H), 2.63 m, 1H), 2.41 (m, 2H), 2.26 - 2.12 (m, 2H), 1.88 (dd, 1H), 1.79 (m, 1H), 1.56 (m, 3H). 1.41 (m, 3H), 1.25 (m, 2H), 1.17 (t, 2H), 1.06 (s, 9H). LRMS (ESI) m / z 714 [(M + H)<sup>+</sup>; izrač.zaC<sub>3</sub>5H4<sub>X</sub>N5O<sub>9</sub>S: 714].
51101 Β
EXAMPLE 15 (5R, 7S, 10S) -10-tert-Butyl-N - ((1R, 2SM-Cyclopropylsulfonyl) amino] carbonyl-2-vinylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo-1,6 , 7,9,10,11,12,14,15,16,17,18dodecahydro-5H-2,22: 5,8-dimethano-4,1,2,2,8,1 l-benzodioxatriazacycloicozin-7-carboxamide (III -206)
<img file="RS51101B_D0091.tif" />
The title compound was prepared according to the procedure used for EXAMPLE 14 (using steps 2 and 3) with the difference that methyl (5R, 7S, 10S) -1-t-butyl-1,15-dimethyl-3,9,12 -trioxo-1,6,7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8-dimethano-
4,13,2,8,1 l-Benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 3, Step 1) used instead of methyl (5R, 7S.10S) -10- tert-butyl-3,9,12-trioxo-1.6, 7,9,10.11,12,14,15,16-dhehydro511-2.22: 5,8-dimethano-4.13.2,8,1 l-benzodioxatriazacycloicosine-7-carboxylate in Step 2. 1 H NMR (400 MHz, ppm , CDC1<sub>3</sub>) b 9.91 (s, 1H). 7.22 (t, 11), 7.09 (d, 2H), 7.05 (d, 1H). 5.77 (m, 1H), 5.60 (s, 1H), 5.45 (d, 1H), 5.29 (s, 1H), 5.15 (d, 1H), 4.72 (q, 2H), 4.40- 4.55 (m, 4H), 4.30 (d, 1H), 4.25 (d, 1H), 3.78 (dd, 1H), 3.26 (d, 1H), 2.91 (m, 1H). 2.50 (m, 3 H), 2.39 (m, 3 H), 2.11 (m, 1 H), 1.98 (m, 2 H), 1.51 (m, 2 H), 1.38 (m, 4 H), 1.18 m, 1H), 1.04 (s, 9H), 1.01 (t, 3H), 0.79 (s, 3H). LRMS (ESI) m / z 742 [(M + H)<sup>+</sup>; calc. for C37II52N5O9S: 742].
51101Β
EXAMPLE 16 (5R, 7S, 10S) -10- (Grc-Butyl-N - ((1R, 2R) -1-Icyclopropylsulfonyl) aminocarbonyl-2-ethylcyclopropyl) T3,9,12-trioxo-1,7,7,9 10, n, 12,14,15,16-decahydro-5H-2,22: 5,8-dimethano4,13,2,8,1 l-benzodioxatriazacyclocycosine-7-carboxamide (W-16)
<img file="RS51101B_D0092.tif" />
<img file="RS51101B_D0093.tif" />
To a solution of methyl (5R, 7S, 10S) -10- [w-butyl-3,9,12-trioxo-1,6,7,9,10,11.12,14,15,16dcahydro-5H-2,22: 5,8-dimethano-4,1,2,2,1,1-benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 14, Step 1) (60 mg, 0.12 mmol) in THF (1 mL) and MeOH (0.5 mL) were added is LiOH (1N 1.17 mL, 1.17 mmol). The reaction mixture was heated to 40 ° C and stirred for 1 hour. when complete consumption of methyl ester starting material was recorded by LC-MS. The mixture was then treated with 0.5 N HCl and EtOAc. The organic layer was then dried over K2CO3, and the solvent was removed in vacuo. The crude product was poured into DMF (1 mL).
To the above solution was added (1R, 2R) -1 - [[(cyclopropylsulfonyl) amino] carbonyl} -2-ethylcyclopropanaminium chloride (32 mg, 0.12 mmol), TBTU (48 mg, 0.15 mmol) and DIPEA (0.044 mL, 0.25 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was purified by reverse phase HPLC to give (5R, 7S, 10S) -10 [m'-butyl'A - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino] carbonyl] -2-ethylcyclopropyl ) -3,9,12trioxo-1,6,7,9,10,1 l, 12,14,15,16-decahydro-5H-2.22: 5,8-dimelano-4,13,2,8,1 l -benzodioxatriazacycloikosin- 7-carboxamide (55 mg, 67% yield). <sup>!</sup>1 H NMR (500 MHz, ppm, CD 3 OD b 7.33 (d, 1H), 7.26 (t, 1H), 7.16 (d, 1H), 6.39 (d, J = 15.7 Hz, 1H), 6.13 (m 1 H), 5.37 (s, 1 H), 4.69 (m, 4 H), 4.47 - 4.28 (m, 4 H), 3.89 (m, 1 H), 3.83 (d, 1 H), 2.98 (m, 1 H) 2.40 (m, 2 H), 2.31 (m, 1 H), 2.11 (t, 1 H), 1.99 (s, 1 H), 1.73 (s, 1 H), 1.60 (m, 2 H) ), 1.52 (m, 1H), 1.29-1.15 (m, 3H), 1.08 (s, 9H), 0.98 (t, 3H), LRMS (ESI) m / z 714 [(M + H)<sup>+</sup>; calc. zaC<sub>3</sub>5H48N<sub>5</sub>O9S: 714].
51101 Β
EXAMPLE 17 (SR, 7SJ0S) -10-tert-Butyl-N4 (1R (2R) -1- {Cyclopropylsulfonyl) amino] carbonyl] -2-ethylcyclopropyl) -3A12-trioxo-6,7,9L0D 1,12,14,15,16 , 17-decahydro-1H, 5H-2,23: 5,8dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosine-7-carboxamide (W-207)
<img file="RS51101B_D0094.tif" />
W-207
Step 1: Methyl (5V, 78,108) -10-GsrH-3,9,12-1pok8o-6,7,9,10,11,12,14,15,16,17decahydro-1H, 5H-2, 23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosin-7-carboxylate
<img file="RS51101B_D0095.tif" />
Methyl (5R, 7S, 10S) -10-Zerc-butyl-3,9,12-trioxo-6,7,9,10,11,12,14,15,16,17-decahydrolH, 5H-2.23: 5 , 8-dimethano-4,13,2,8,11-benzodioxazazacyclohenicosine-7-carboxylate was prepared according to the procedure used for methyl (5R, 7S.10S) -1O- [m'-butyl-15,15-dimethyl- 3,9,12-trioxo-1,6,7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8-dimethano-
4,13,2,8,1 1-Benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 3, Step 8) except that 3-methyl-N - [(hex-5-enyloxy) carbonyl] -L-valine according to the procedure given below) was used instead of A - [[(2,2-dimethylpent-4-enyl) oxy] carbonyl} -3methyl-L-valm in Step 7. LRMS (ESI) m / z 528 [(M + H)<sup>+</sup>; calc. for C28H38N3O7: 528].
51101 Β
Step 2: (5R, 7SJ0S) -10-yl> Butyl-N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-ethylcyclopropyl) -3,9,12-trioxo-6 , 7,9,10,1,11,1,1,14,15,16,17-decahydro-1H, 5H2,23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosin-7-carboxamide
EXAMPLE 17 was prepared according to the procedure used for EXAMPLE 16 with the difference that methyl (5R, 7S.10S) -10- [erc-butyl-3,9,12-trioxO ”was used.
6,7,9,10,11,12,14,15,16,17-decahydro-1H, 5H-2,23: 5,8-dimethano-4,1,2,2,8,11benzodioxatriazacyclohenicosine-7-carboxylate instead of methyl- ( 5R, 7S, 10S) -10- [tert-butyl-
3,9,12-trioxo-1,6,7,9,10,11,11,15,16-decahydro-5H-2,22: 5,8-dimethano-4,13,2,8 , 11-benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 14. Step 1). 1 H NMR (500 MHz, ppm, CD<sub>3</sub>OD) b 9.06 (s, 1H), 7.27 (t, 1H). 7.24 (d, 1H), 7.18 (d, 1H), 6.40 (d, J = 16.4 Hz, 1H), 6.11 (m, 1H), 5.39 (t, 1H), 4.80 (d, 1H) ). 4.69 (m, 4H). 4.42 (s, 1H), 4.25 (d, 1H). 3.97 (dd, 1H), 3.79 (quin., 1H), 2.98 (m, 1H), 2.50 (q, 1H), 2.78 (m, 2H), 2.15 (m, 1H). 1.77 - 1.54 (m, 8 H), 1.32 - 1.19 (m, 4 H), 1.11 (m, 1 H), 1.07 (s, 9 H), 0.98 (t, 3 H). LRMS (ESI) mtz 728 [(M + H))<sup>+</sup>; calc. for C36H5ON5O9S: 728].
EXAMPLE 18 (5R, 7S, 10S) -10- (Grc-Butyl-N - ((1R, 2R) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-ethylcyclopropyl) -15,15-dimethyl-3,9, 12-trioxo-6,7,9,10,11,11,14J5,16,17-decahydro-1H, 5H2,23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosin-7- carboxamide (W-208)
<img file="RS51101B_D0096.tif" />
<img file="RS51101B_D0097.tif" />
51101 Β
Step 1: Methyl (5K, 78,108) -10- / egs-acid ~ 15,15- {Ip1ep1 ~ 3,9,12-Mok5o6,7,9,10,11l, 12,14,15,16,17- decahydro-1H, 5Ii-2,23: 5,8-dimethano-4,13,2,8,1,1-benzodioxatriazacyclohenicosine-7-carboxylate
<img file="RS51101B_D0098.tif" />
Methyl (5R, 7S, 10S) -10- [tert-butyl-15,15-dimethyl-3,9,12-trioxo-6,7,9,10,11,12,14,15,
16.17-decahydro-1H, 5H-2,23: 5,8-dimethano-4,13,2,8,1 l-benzodioxatriazacyclohenicosin-7-carboxylate was prepared according to the procedure used for methyl (5R, 7S, 10S) -10 -fert-butyl15,15-dimethyl-3,9,12-trioxo-1,6,7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8-dimethano-
4,13,2,8,1 l-benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 3, Step 8) except that X - {[(2,2-dimethylhex-5-enyl) oxy] carbonyl} -3- methyl-L-valine (prepared according to the procedure given below) used instead of A '- {[(2,2-dimethylpent-4-enyl) oxy] carbonyl} -3-methyl-L-valine in Step 7. LRMS ( ESI) m / z 556 [(M + H)]<sup>+</sup>; izraČ. for C30H42N3O7: 556]. Step 2: (5R, 7S, 10S) -10- / m'-Butyl-A<sup>,</sup>- ((1R, 2R) -1 - {(cyclopropylsulfonyl) amino] carbonyl} -2-ethylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo-6,7,9,10,11.12,14 , 15,16,17decahydro-1H, 5H-2,23: 5,8-dimethano-4,1,2,2,8,11-benzodioxatriazacyclohenicosin-7-carboxamide
EXAMPLE 18 was prepared according to the procedure used for EXAMPLE 16 with the difference that methyl (5R, 7S, 10S) -10-tor-butyl-15,15-dimethyl-3,9,12-trioxo-
6.7.9.10.11.12.14.15.16.17-decahydro-1H, 5H-2,23: 5,8-dimethano-4.13,2,8,1-1-benzodioxatriazacyclohenicosine-7-carboxylate instead of methyl- (5R, 75.10S) - 10-to-t-butyl 3,9,12-trioxo-1,6,7,9,10,11,12,14,15,16-decahydro-5H-2,22: 5,8-dimethano-4 , 13,2,8,11 benzodioxatriazacycloicosine-7-carboxylate (EXAMPLE 14, Step 1). 1 H NMR (500 MHz, ppm, CD 3 OD) δ 10.05 (s, 1H). 7.24 (m, 2H), 7.17 (d, 1H). 7.11 (d, 1H), 6.61 (s, 1H). 6.28 (d, 16.4 Hz, 1H), 5.95 (m, 1H), 5.58 (m, 1H), 5.31 (s, 1H), 4.71 (m, 2H), 4.55 (m, 2H) ,,
4.46 (d, 2H), 4.29 (dd, 1H), 4.17 (d, 1H). 3.89 (d, 1H), 3.32 (d, 1H). 2.92 (m, 1H). 2.59 (m, 1H), 2.21-2.30 (m, 2H), 2.08 (m, 1H), 1.60-1.78 (m, 6H), 1.22-1.31 (m, 5H). 1.06 (p. 9
51101 Β
Η), 1.04 (ί, 3 Η), 0.093 (t, 3 (), 0.87 (s, 3 Η). LRMS (ESI) m / z 756 [(Μ + Η)<sup>+</sup>; calc. for
C3SH54N5O9S: 756],
Preparation / V4 (Pent-4-eA-yloxy) carbonyl] -L-norleucine:
ΌΗ
To a solution of 1-penten-4-ol (0.95 g, 11.0 mmol) 11 DMF (15 mL) at 0 ° C was added carbonyldiimidazole (1.79 g, 11.0 mmol). The reaction mixture was warmed to room temperature and stirred for 30 min. Then L-norleucine methyl ester hydrochloride (2.0 g, 11.0 mmol) was added, the reaction mixture was heated to 50 ° C and stirred for 15 min. After cooling, the reaction mixture was diluted with ethyl ether and washed twice with water. The organic layer was dried over sodium sulfate, filtered and concentrated. The crude product was purified by chromatography on silica gel (eluting with a gradient of 10 to 90% ethyl acetate in hexanes) to give 2.1 g (74% yield) of methyl A - [(pent-4-en-1-yloxy) carbonyl] - L-norleucinate as a clear oil.
In a mixed solution of methyl A<sup>r</sup>- [(pent-4-enyloxy) carbonyl] -L-noryucinate (8.50 g, 33.03 mmol) in THF (20 mL) was added to 1N NaOH (20 mL). This reaction solution was stirred at room temperature for 3 hours, then acidified to pH 3 with 1N HCl and extracted with (3 x 250 μm) EtOAc. The combined EtOAc layer was washed with 50 mL of water, 50 μL of brine, dried over sodium sulfate, filtered and concentrated to give 7.09 g (88% yield) of the title product as a clear oil. LRMS (ESI) m / z 244 [(M + H)<sup>+</sup>; calc. for C12H22NO4: 244],
Preparation of 3-Methyl-A - [(pent-4-cynyloxy) carbonyl-L-valine:
A solution of 4-pentenol (7.22 g, 83.8 mmol) and triphosgene (11.3 g, 38.1 mmol) in dioxane (160 mL) was cooled to 0 ° C and then DIPEA (9.85 g, 76.2 mL) was added dropwise. The white suspension was stirred vigorously for 1 hour at 25 ° C, then cooled to 0 ° C. 1 N NaOH solution (76.2 mL) and t-butylglycine (10.0 g, 76.2 mmol) were added. The resulting suspension was heated to 25 ° C and stirred for 18 hours. Approximately half of the dioxane was removed in vacuo. solution poured
51101Β in 1 N NaOH (100 mL) and washed with dichloromethane (3 h 150 mL). The aqueous layer was acidified with 6 N HCl and the desired product was extracted with dichloromethane (3 x 150 niL). The combined organics were dried over MgSO 4<sub>4</sub> and concentrated to give 13.7 g (73.9% yield) of 3-methyl-N - [(pent-4-enyloxy) carbonyl] -L'Valine as a colorless oil. LRMS (ESI) m / z 244 [(M + M & lt; + & gt;)<sup>+</sup>; calc. for C12H22NO4: 244].
Preparation of N-Hex-5-en-1-yloxy) carbonyl-1-noryucine:
<img file="RS51101B_D0099.tif" />
N - [(Hex-5-en-1-yloxy) carbonyl] -L-norleucine was prepared according to the procedure for N [(pent-4-en-1-yloxy) carbonyl] -L-norleucine using 5-hexenol instead of 4 -pentenol. LRMS (ESI) m / z 258 [(M + H) & lt; + & gt;]<sup>+</sup>; calc. for C<sub>13</sub>H<sub>24</sub>NO<sub>4</sub>: 258],
Preparation of 3-Methyl-N - [(hex-5-enyloxy) carbonyl | -L-valine:
<img file="RS51101B_D0100.tif" />
3-Methyl-N - [(hex-5-enyloxy) carbonyl] -L-valine was prepared according to the procedure for 3-methyl-A - [(pent-4-enyloxy) carbonyl] -L-valine using 5-hexenol instead of 4- pentenol. LRMS (ESI) m / z 258 [(M + H) & lt; + & gt;]<sup>+</sup>; calc. for C<sub>13</sub>H<sub>24</sub>NO<sub>4</sub>: 258].
Preparation of N- [Hept-6-en-1-yloxy) carbonyl-L-norleucine:
<img file="RS51101B_D0101.tif" />
A - [(Hept-6-en-1-yloxy) carbonyl] -L-norleucine was prepared according to the procedure for N [(pent-4-en-1-yloxy) carbonyl] -L-norleucine using 6-heplenol innesto 4 -pcntcnola. LRMS (ESI) mtz 272 [(M + H))<sup>+</sup>; calc. for C<sub>14</sub>H<sub>26</sub>NO<sub>4</sub>: 272],
51101Β
Preparation of α-[[(2,2-Dimethylpent-4-enyl) oxylcarbonyl) -3-methyl-L-valine:
<img file="RS51101B_D0102.tif" />
Step 1: 2,2-Dimethylpent-4-en-1-ol
A solution of 2,2-dimethyl allyl acetic acid (6.0 g, 46.8 mmol) in anhydrous THF was cooled in an ice bath to 0 ° C. A light stream of 1M Lithium aluminum hydride in THF (56.2 mL, 56.2 mmol) was added and the reaction was allowed to warm to 25 ° C. The reaction mixture was stirred for 1 hour before pouring into 1N HCl and diethyl ether. The organic layer was separated, dried over MgSO 4<sub>4 </sub>and concentrated to give 2,2-dimethylpent-4-en-1-ol as a clear oil (4.7 g, 87.9% yield).
Step 2: N - {[(2,2-Dimethylpent-4-phenyl) oxycarbonyl} -3-methyl-L-valine
DIPEA (2.48 g, 19.2 mmol) was added dropwise to 0 ° C a solution of 2,2-dimethylpent-4en-1-ol (2.24 g, 19.6 mmol) and triphosgene (2.56 g, 8.64 mmol) in 60 nL dioxane. The resulting white suspension was stirred at 0 ° C for 5 min, then allowed to warm to 25 ° C over 1 h. The suspension was cooled to 0 ° C with an ice bath, followed by the addition of 1 N NaOH (19.2 mL) and 7-to-c-butylglycine (2.52 g, 19.2 mmol). The reaction mixture was heated to 25 ° C and stirred for 72 hours. Dioxane was removed in vacuo and the reaction mixture was basified to pH 12 with 1 N NaOH. The aqueous layer was extracted with dichloromethane (Zh 150 mL), then acidified to pH 11 with 6 N HCl. The aqueous layer was extracted with dichloromethane (3 x 150 mL). The combined organic layers were dried over MgSO 4<sub>4</sub> and concentrated to give L - {[(2,2-dimethylpent-4-enyl) oxy] carbonyl-3-methyl-L-valine as a white powder (4.26 g, 827% yield). LRMS (ESI) m / z 272 [(M + H) & lt; + & gt ;.<sup>+</sup>; calc. for C,<sub>4</sub>Hz<sub>6</sub>NO<sub>4</sub>: 272],
51101Β
Preparation of α-β (2,2-Dimethylhex-5-enyl) oxy] carbonyl} -3-methyl-L-valine:
Step 1: Ethyl 2,2-dimethylhex-5-enoate
<img file="RS51101B_D0103.tif" />
To a stirred solution of diisopropylamine (13.38 mL, 94.70 mmol) in anhydrous THF (50 mL), at -70 ° C and under nitrogen, was slowly added 2.5 M n-BuLi in ether (36.50 mL, 91.25 mmol). The solution was stirred for 15 minutes, ethyl isobutyrate (11.51 mL, 86.09 mmol) in THF (50 mL) was then added dropwise to this reaction solution, and the solution was stirred for 20 minutes before 4-bromo-1-butene was added dropwise (9.79 mL ·, 96.42 mmol) in HMPA (20 mL). The reaction solution was then stirred to -50 ° C for 5 hours, quenched with 1M IICl (10 mL) and water (100 mL), then extracted with (3 x 125 mL) ether. The combined ether layer was washed with aqueous (4 x 70 mL), aqueous saturated NaHCO 3 solution.<sub>3</sub> (2 x 70 mL), dried over Na<sub>2</sub>SO<sub>4</sub>, filtered and concentrated. The crude product was subjected to ilash chromatography on 120 g of silica gel 60, eluting with 1-20% EtOAc / hexanes to give the title compound as a clear oil (1 l.Olg, 75% yield). LRMS (ESI) m / s 171 [M + II]<sup>+</sup>: calc. for CjoIIi-A: 171],
Step 2: 2,2-Dimethylhex-5-en-1-ol
To a stirred solution of 1M LAH in ether (142.14 mL, 142.14 mmol), at 0 ° C and under nitrogen, ethyl 2,2-dimethylhex-5-enoate (11.00 g, 64.61 mmol) dissolved in 100 mL of anhydrous ctra was added dropwise over 1 hour. This reaction solution was stirred at 22 ° C for 20 hours. then quenched with water (3 mL), 1M NaOH (11 mL) and water (9 mL), dried over Na<sub>2</sub>SO<sub>4</sub>, filtered and concentrated to give the title product (7.22 g, 87.09%). 1 H NMR (500
51101 Β
ΜΗζ, CDCl 3) δ 5.85-5.77 (m, 1H); 5.01 (d, 1H); 4.93 (d. 1 (): 3.33 (d, 2II); 2.03 (m, 2H); 1.34 (m, 2H); 0.89 (m, 6H) ppm.
KogakZ: A - {[(2,2-Dimethylhex-5-enyl) oxy] carbonyl} -3-methyl-L-valine
To a stirred solution of 2,2-dimethylhex-5-en-1-ol (10.75 g, 83.85 mmol) in anhydrous 1,4-dioxane (100 mL), at 0 ° C and under nitrogen, was added triphosgene (13.69 g, 46.12 mmol) followed by DIPEA (14.61 mL, 83.85 mmol) carefully. This reaction solution was stirred at 22 ° C for 1 h, cooled to 0 ° C and 1N NaOH (83.85 mL, 83.85 mmol) and L- / erc-leucine (11.00 g, 83.85 mmol) were added slowly, then stirred at 22 ° C. ° C for 20 hours. The reaction solution was basified to pH 10 with 1N NaOH, washed with SNJSE (Zh 100 mL), acidified to pH 5 with 1N HCl and extracted with CII2Cl2 (3 x 150 mL). The combined CH2Cl2 layer was washed with water (100 mL). dried over Na 2 SO 4 filtered and concentrated to give the title product (20.26 g, 84.66%). 1 H NMR (500 MHz, CDCl 3)<sub>3</sub>b 5.85-5.77 (m, 1H): 5.24 (d, 1H); 5.01 (d, 1H); 4.93 (d, 1H); 4.20 (d, 1H); 3.86 (d, 1H); 3.79 (d, 1H); 2.01 (m, 2 14): 1.36 (m, 2H); 1.04 (s, 9H); 0.92 (m, 6H) ppm. LRMS (ESI) m / z 286 [(M + N)<sup>+</sup>: calc. for 286],
Preparation of (1R, 2R-1-yl (cycloprodibulfonyl) amino] carbonyl) -2-ethylcyclopropanaminium chloride:
<img file="RS51101B_D0104.tif" />
Mixture (1A, 2<sub>1</sub>S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropanaminium chloride (Llinas-Brunet et al. US03 / 15755 and Wang et al. WO 03/099274) (0.05 g, 0.187 mmol) and palladium on carbon (10 % w / w (0.0lg) in EtOAc (5 mL) was stirred vigorously under a hydrogen atmosphere with a hydrogen balloon for 1 hour. The reaction mixture was filtered and concentrated to give (17? .27?) - 1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2ethicyclopropanaminium chloride (0.045 g, 89% yield).
51101Β
EXAMPLE 19 (5R, 75,105) -10H-Butyl-N - ((1R) 25) -1 - {[(cyclopropylsulfonyl) amino] carbonyl]<sub>?</sub>2-vinylcyclopropyl) -15,15-dimethyl-3,9,12-trioxo-6,7,9,10,11,1,1,14,15,16,17,18,19dodecahydro-1, 7, 5 # -2 , 23: 5,8-diniethano-4,13,2,8,11-benzodioxatriazacyc] ohenicosin-7-carboxamide (W-210)
<img file="RS51101B_D0105.tif" />
EXAMPLE 19 was prepared from (57?, 75,105) -10- [tert-butyl-15,15-dimethyl-3,9,12-trioxo6,7,9,10,11,12,14,15,16,17 , 18,19-dodecahydro-177,577-2,23: 5,8-dimethano-4,13,2,8,11 benzodioxatriazacyclohenicosine-7-carboxylic acid (EXAMPLE 13 Alternative Preparation, Kogak 4) using the procedure for EXAMPLE 3, Kogak 10 n NMR (500 MHz, CD 3 OD, ppm) δ 7.25-7.09 (m, 3 H). 5.82-5.74 (m, 1H), 5.35-5.29 (m, 2H), 5.15-5.12 (m, 1H), 4.75-4.59 (m, 3H). 4.45-4.38 (m, 2H), 4.21-4.12 (m, 1H), 4.13-4.09 (m, 1H), 3.95-3.92 (m, 1H). 2.982.94 (m, 1H), 2.62-2.54 (m, 1H), 2.49-2.46 (m, 2H), 2.25-2.21 (m, 1H), 2.19-2.13 (m, 1H), 1.90-1.88 (m, 1H), 1.52 (m, 2H), 1.48-1.45 (m, 1H), 1.40-1.18 (m, 6H), 1.15-1.00 (m, 14H), and 0.81 (m, 4H). LRMS (ESI) m / z 756.4 [(M + H)]<sup>+</sup>; calc. for SzzNzz ^ SŽ 755.9],
51101 Β
EXAMPLE 20 (51?, 75,105 ', 18 £) -10-Cyclohexyl-1<sup>)</sup>V - ((ll?, 2<sub>1</sub>y) -1n (cyclopropylsulfonyl) amino carbonyl) -2-vinylcyclopropyl-15,15-dimethyl-3,9,12-trioxo-6,7,9,10,11,1,1,14,15,16,17decahydro-1H, 5H-2, 23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohenicosin-7-carboxamide (W-225)
<img file="RS51101B_D0106.tif" />
EXAMPLE 20 was prepared using the procedures of EXAMPLE 13 Alternative Preparation, Steps 1, 2, 4 and 5 using (2X) -cyclohexyl [[[(2,2-dimethylhex-5-en-yl) oxy] carbonyl] amino) acetic acid in Step 1 and (1R, 2S) -1 - [[(cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropanaminium chloride in Step 5. 1 N NMR (500 MHz. CD<sub>3</sub>OD, ppm) b 7.26 (m, 1H), 7.20 (t, J = 7.5 Hz, 1H), 7.15 (d, J = 9.5 Hz, 1H). 6.38 (d, .7 = 9.5 Hz, 1H), 5.99-6.02 (m, 1H), 5.74-5.80 (m, 1H), 5.29-5.34 (m, 2H), 5.11-5.14 (m, 1 H), 4.794.81 (m, 2 H), 4.64-4.72 (m, 3 H), 4.56 (d, J = 11.5 Hz, 1 H), 4.36-4.40 (m, 2 H), 4.18 (d J = 11.5 Hz, 1 H), 4.10 (d, J = 5.5 Hz, 0.5 H), 3.91-3.94 (dd, .7 = 11.5, 3.5 Hz, 1 H), 3.34 (d, J = 11.0 Hz, 1H), 2.95-2.97 (m, 1H), 2.52-2.56 (m, 1H), 2.16-2.35 (m, 5H), 1.65-1.82 (m, 8H), and 0.85-1.43 (m , 17 H). LRMS (ESI) m / z 780.4 [(M + H)<sup>+</sup>: calc. for SzoN ^ S ^ b: 780.9].
51101Β
EXAMPLE 21 (5R, 75'.l ()<sub>t</sub>V) -10-Cyclohexyl-N - ((R, 2R) -1H (cyclopropylsulfinyl) amino) carbonyl (nyl} 2-ethylcyclopropyl) -155-dimethyl-3,9J2-trioxo-6, 7,9J0,11,11,1,14,15,16,17,18,19dodecahydro-ΙΗ, 57 / -2,23: 5,8-dimethano-4,13,2,8,11-benzodioxatriazacyclohecyloicin-7-carboxamide (III -226)
<img file="RS51101B_D0107.tif" />
EXAMPLE 21 was prepared from EXAMPLE 20 using the procedure described for EXAMPLE 8. 1 H NMR (500 MHz, CDCl 3)<sub>3</sub>, ppm) b 10.13 (s, 1 H), 7.22 (t. J = 7.5 Hz, 1 H), 7.10 (d, ./=7.5 Hz, 1 H), 7.05 (d. J = 7.5 Hz, 1 H) ), 6.73 (s, 1H), 5.40 (d, J = 9.5 Hz, 1H), 5.36 (m, 1H), 4.67-4.76 (m, 2H), 4.55 (d, 15.5 Hz, 1H) ), 4.44 (d, 14.5 Hz, 1H), 4.41 (d, J = 11.0 Hz, 1H), 4.29-4.39 (m, 2H). 4.16 (d, J = 11.0 Hz, 1H), 3.82-3.85 (dd, J = 11.5,3.5 Hz, 1H), 3.25 (d, J = 11.0 Hz, 1H), 2.95 (m, 1H) , 2.51-2.59 (m, 2H), 2.36-2.44 (m, 2H). 1.73-1.76 (m, 5H) and 0.79 (br s, 2H). LRMS (ESI) m / z 784.4 [(M + H)]<sup>+</sup>; calc. for C40H57N5O9S: 784.4],
Alternative preparation (1S, 2R) -1-amino-N '- (cyclopropylsulfonyl) -2-ethylcyclopropane carboxamide hydrochloride:
HCI
<img file="RS51101B_D0108.tif" />
51101Β
Step 1: tert-Butyl (1R, 2R) -1 {[(cyclopropylsulfonyl) amino] carbonyl} -2-ethylcyclopropyl) carbamate:
The hydrogenation apparatus was charged with a methanological (1000 mL) suspension of tert-butyl (1R, 2S) -1 - {[(cyclopropylsulfonyl) amino] carbonyl} -2-vinylcyclopropyl) carbamate (164 g, 0.50 mol) (Wang et al, US 6,995,174) and 5% Ru / C (dry, 7.5% by weight, 12.4 g) and adjusted for mixing. The apparatus was placed under nitrogen (20 psig) and ventilated under atmospheric pressure three times to remove residual oxygen. The apparatus was then placed under hydrogen (50 psig). After 20 hours, the apparatus was ventilated under atmospheric pressure. The reaction suspension was then transferred out of reaction and filtered through brine (34 grams, wet weight / ΙΟΟ mL methanol) to give a clear, light brown solution. The solka flock was washed with methanol (200 mL h 2). The combined methanol solutions were concentrated under reduced pressure to give the crude product as a white solid (153 g). The crude product was suspended in ethyl acetate (800 mL), heated to 40 ° C and allowed to age for 30 minutes. The solution was then seeded, allowed to age for 30 minutes, and heptane (500 mL) was added via addition funnel over 30 minutes. The partially crystallized solid was cooled to room temperature and allowed to age overnight after which a new amount of heptane (500 mL) was added. After an hour. a new amount of heptane (250 mL) was added via a addition funnel, and the white suspension was allowed to age for one hour. The solution was filtered and the solid was washed with heptane / EtOAc (500 mL, 4: 1) and dried under reduced pressure to give tert-butyl (1R, 2R) -1 - ([(cyclopropylsulfonyl) amino] carbonyl) -2-ethylcyclopropyl) carbamate (125.9 g).
Step 2: (1R, 2R) -1-Amino-N- (cyclopropylsulfonyl) -2-ethylcyclopropanecarboxamide hydrochloride:
A solution of the product from Step 1 above (92 g, 0.28 tnol) in DCM (1200 mL) was cooled to 0 ° C and HCl was passed through the solution for 10 min. the cooling bath was removed and the reaction mixture was stirred for 2 hours. Nitrogen was passed through the reaction
51101 Β The mixture was evaporated for 5 minutes and the volatiles were evaporated. The residue was azeotroped with DCM (x 3) to give an off-white powder (75 g). LRMS (M + H)<sup>+</sup> calc. = 233; found 233.
Preparation of _______ (25) -cyclohexyl ({2,2-dimethylhex-5-en-1-yl) oxy] carbonyl] amino) acetic acid:
<img file="RS51101B_D0109.tif" />
(2S-Cyclohexyl ({[(2,2-dimethylhex-5-en-1-yl) oxy] carbonyl} amino) acetic acid was prepared according to the procedure for 3-methyl-N - [(pent-4-cynyloxy) carbonyl-1-valine using (2S) -amino (cyclohexyl) acetic acid and 2,2-dimethylhex-5-en-1-ol LRMS (ESI) m / z 312.3 [(M + H)]<sup>+</sup>; calc. for C<sub>17</sub>H<sub>30</sub>NO<sub>4</sub>: 312.2],
EXAMPLE 22
Time-decomposed fluorescent assay (TRF) of HCV NS3 protease
The TRF test of NS3 protease was performed in a final volume of 100 μΐ in test buffer containing 50 mM HEPES, pH 7.5, 150 mM NaCl, 15% glycerol, 0.15% Triton H-100, 10 mM DTT and 0.1% PEG 8000. NS3 protease was pre-incubated with different concentrations of inhibitor for 10-30 minutes. The peptide substrate for the test is Ac-C (Eu) -DDMEE-Abu [COO] -XSAK. (QSY7) -NH2, where the Eu group is labeled with europium, Abu is a 1-aminobulemic acid that binds an ester bond to 2-hydroxy propionic acid (X). Hydrolysis of the peptide via NS3 protease activity causes the fluorophore to separate from the luminescence quenching material, resulting in increased fluorescence. Protease activity was initiated by the addition of TRF peptide substrate (final concentration 50-100 nM). The reaction was quenched after 1 h at room temperature with 100 μΐ 500 mM MES, pH 5.5. Fluorescence of the product was detected using either a Victor V2 or a fusion fluorimeter (Perkin Elmer Life and Analytical Sciences) with excitation at 340 nm and emission at 615 nm with a delay of 50400 ps. The tested concentrations of different enzyme forms were selected with a signal ratio
51101 Β against a background of 10-30. Inhibition constants were derived using fitting based on four parameters.
The compounds in Examples 1-21 were tested to have a Ki value of less than 100 nM (e.g., less than 1 nM) in the TRF assay of NS3 protease as described above.
Contents25
109 sheets
Sheet 1 Sheet 2 Sheet 3 Sheet 4 Sheet 5 Sheet 6 Sheet 7 Sheet 8 Sheet 9 Sheet 10 Sheet 11 Sheet 12 Sheet 13 Sheet 14 Sheet 15 Sheet 16 Sheet 17 Sheet 18 Sheet 19 Sheet 20 Sheet 21 Sheet 22 Sheet 23 Sheet 24 Sheet 25 Sheet 26 Sheet 27 Sheet 28 Sheet 29 Sheet 30 Sheet 31 Sheet 32 Sheet 33 Sheet 34 Sheet 35 Sheet 36 Sheet 37 Sheet 38 Sheet 39 Sheet 40 Sheet 41 Sheet 42 Sheet 43 Sheet 44 Sheet 45 Sheet 46 Sheet 47 Sheet 48 Sheet 49 Sheet 50 Sheet 51 Sheet 52 Sheet 53 Sheet 54 Sheet 55 Sheet 56 Sheet 57 Sheet 58 Sheet 59 Sheet 60 Sheet 61 Sheet 62 Sheet 63 Sheet 64 Sheet 65 Sheet 66 Sheet 67 Sheet 68 Sheet 69 Sheet 70 Sheet 71 Sheet 72 Sheet 73 Sheet 74 Sheet 75 Sheet 76 Sheet 77 Sheet 78 Sheet 79 Sheet 80 Sheet 81 Sheet 82 Sheet 83 Sheet 84 Sheet 85 Sheet 86 Sheet 87 Sheet 88 Sheet 89 Sheet 90 Sheet 91 Sheet 92 Sheet 93 Sheet 94 Sheet 95 Sheet 96 Sheet 97 Sheet 98 Sheet 99 Sheet 100 Sheet 101 Sheet 102 Sheet 103 Sheet 104 Sheet 105 Sheet 106 Sheet 107 Sheet 108 Sheet 109
57 members in 34 offices
Priority claims12
| Document | Office | Kind | Date |
|---|---|---|---|
| 70076405 | United States of America | P | |
| 70076405 | United States of America | P | |
| 72456605 | United States of America | P | |
| 72456605 | United States of America | P | |
| 2006027573 | United States of America | W | |
| 2006027573 | United States of America | W | |
| 20050700764P | – | – | – |
| 20050724566P | – | – | – |
| PCTUS2006027573 | – | – | – |
| US20050700764P | – | – | – |
| US20050724566P | – | – | – |
| WO2006US27573 | – | – | – |
Members57
| Document | Office | Kind | |
|---|---|---|---|
| US2007027071A1 | United States of America | A1 | |
| AU2006276189A1 | Australia | A1 | |
| CA2615022A1 | Canada | A1 | |
| WO2007015787A1 | World Intellectual Property Organization (WIPO) | A1 | |
| WO2007015855A1 | World Intellectual Property Organization (WIPO) | A1 | |
| TW200740850A | Taiwan Province of China | A | |
| AR057456A1 | Argentina | A1 | |
| CR9621A | Costa Rica | A | |
| MX2008000866A | Mexico | A | |
| KR20080027860A | Republic of Korea | A | |
| IL188618A0 | Israel | A0 | |
| EP1910404A1 | European Patent Office (EPO) | A1 | |
| NO20080879L | Norway | L | |
| EP1924593A1 | European Patent Office (EPO) | A1 | |
| EA200800371A1 | Eurasian Patent Organization (EAPO) | A1 | |
| CN101228181A | China | A | |
| US7470664B2 | United States of America | B2 | |
| JP2009502793A | Japan | A | |
| MA30198B1 | Morocco | B1 | |
| HK1121169A1 | Hong Kong, China | A1 | |
| US2009124661A1 | United States of America | A1 | |
| TNSN08033A1 | Tunisia | A1 | |
| EP1924593B1 | European Patent Office (EPO) | B1 | |
| AT443075T | Austria | T | |
| ATE443075T1 | Austria | T1 | |
| DE602006009280D1 | Germany | D1 | |
| EP1924593B8 | European Patent Office (EPO) | B8 | |
| PT1924593E | Portugal | E | |
| DK1924593T3 | Denmark | T3 | |
| HRP20090627T1 | Croatia | T1 | |
| ES2332821T3 | Spain | T3 | |
| PL1924593T3 | Poland | T3 | |
| EA013331B1 | Eurasian Patent Organization (EAPO) | B1 | |
| SI1924593T1 | Slovenia | T1 | |
| EP1924593B9 | European Patent Office (EPO) | B9 | |
| UA90909C2 | Ukraine | C2 | |
| ZA200800072B | South Africa | B | |
| SG164376A1 | Singapore | A1 | |
| NZ565223A | New Zealand | A | |
| RS51101BThis record | Serbia | B | |
| MY142240A | Malaysia | A | |
| BRPI0613633A2 | Brazil | A2 | |
| JP4621282B2 | Japan | B2 | |
| JP2011079828A | Japan | A | |
| GEP20115248B | Georgia | B | |
| AU2006276189B2 | Australia | B2 | |
| AU2006276189B9 | Australia | B9 | |
| US8216999B2 | United States of America | B2 | |
| CA2615022C | Canada | C | |
| TWI387603B | Taiwan Province of China | B | |
| KR101296095B1 | Republic of Korea | B1 | |
| CN101228181B | China | B | |
| JP5345117B2 | Japan | B2 | |
| EP1910404B1 | European Patent Office (EPO) | B1 | |
| ES2446015T3 | Spain | T3 | |
| EP1910404B9 | European Patent Office (EPO) | B9 | |
| ES2446015T9 | Spain | T9 |
Numbers
- Publication
- 51101
- Publication, DOCDB
- 51101
- Publication, EPODOC
- RS51101
- Application
- 20090548
- Application, DOCDB
- P20090548
- Application, EPODOC
- RS2009P000548
Titles2
- English
- NCV NS3 PROTEASE INHIBITORS
- Serbian
- INHIBITORI HCV NS3 PROTEAZE
Classification
- CPC, 12
- C07K5/0808
- C07D487/18
- A61K38/00
- A61P1/16
- C07K5/0812
- A61P31/00
- C07K5/0827
- A61P31/12
- A61P31/14
- A61P43/00
- C07D487/22
- A61K31/407
- IPC, 3
- C07K5 08
- A61K38 06
- A61P31 14