Methods for synthesizing metal mesoporphyrins

The present application claims priority over Provisional Patent Application No. 61 / 469,791 filed on March 30, 2011, No. 61 / 469,792 filed on March 30, 2011 and No. 61 / 532,301 filed on September 8, 2011, each of them titled "Methods of Mezoporphyrin Metals Synthesis".

C. Government interest: Not applicable

D. Parties to the Joint Research Agreement: Not applicable

E. Inclusion by Reference of the Material included on the Compact Disc: Not applicable

F. Background of the invention

WO 2008/045377 relates to large-scale (batch) compositions containing stannsoporfin of high purity, as well as methods for synthesizing such compositions.

Zvezdina et al. (Russian Journal of Inorganic Chemistry, 2006, Volume 51, No. 1, pp. 112-117) describes a study on cadmium protoporphyrin trans-metallation (CdPP) using zinc chloride and dimethyl sulfoxide (DMSO) and using cobalt chloride in acetonitrile.

G. Summary of the Invention [0001] The embodiments described herein are generally directed to methods for synthesizing tin mesoporphyrin compounds.

[0002] In some aspects, the method for synthesizing a mesoporphyrin compound includes heme transmetalation and hydrogenation of tin protoporphyrin IX. In some embodiments, the heme is trans metalized in the presence of iron sulfate. In some embodiments, the metal protoporphine IX is hydrogenated using dissolved ammonium hydroxide, dimethylformamide or n-methylpyrrolidone. In some embodiments, tin mesoporphyrin is precipitated using methyl tert-butyl ether (MTBE). The metal being introduced is tin. In embodiments, the tin is incorporated into the mesoporphyrin to provide stannsoporfin using tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2-ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate.

[0003] In some aspects, the method of synthesizing a tin mesoporphyrin compound involves forming the protoporphyrin methyl ester from hemin and converting an intermediate of protoporphyrin methyl ester to a tin mesoporphyrin compound by incorporating tin and hydrogenation. In some embodiments, the introduction of tin yields the intermediate product of tin protoporphyrin dimethyl ester. In further embodiments, the tin protoporphyrin dimethyl ester is hydrogenated in dichloromethane on a palladium catalyst to give the tin mesoporphyrin dimethyl ester. In embodiments, the tin mesoporphyrin dimethyl ester is heated in dissolved ammonium hydroxide to give the tin mesoporphyrin compound. The metal being introduced is tin.

H. Description of the Drawings [0004] For a more complete understanding of the nature and advantages of the present invention, reference should be made to the following detailed descriptions in connection with the attached drawings, where:

[0005] Figure 1 depicts an exemplified low-temperature anaerobic synthesis of stannsoporfin and the synthesis of each component.

[0006] Figure 2 shows an exemplary oxidative synthesis of stannsoporfin with reflux.

[0007] Figure 3 shows the structure of stannsoporfin (B992).

[0008] Figure 4 shows the structure of monovinyl intermediates (A) CJ9 and (B) CKO.

[0009] Figure 5 shows the structure of tin protoporphyrin (CH8).

I. Detailed Description [0010] Before describing the present composition and methods, it is obvious that this invention is not limited to a particular method, composition or methodology as they may be subject to change. It is also evident that the terminology used in the description is given only for describing particular versions or embodiments and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Preferred materials and methods are described herein, although any methods, devices and materials similar or equivalent to those described herein, may be used to apply or test this embodiment of the present invention. Nothing in the present description can be considered as an admission that the inventors are not entitled to anticipate such disclosure in favor of the earlier invention.

[0011] It should also be noted that throughout this document as well as in the appended claims, singular forms also include the plural unless the context clearly dictates otherwise. For example, reference to "a compound" means thus a reference to one or more compounds and includes equivalents known to those skilled in the art and the like.

[0012] As used herein, the term "about" means plus or minus 10% of the numerical value of the number that is used. Thus, about 50% means a range of 45% -55%.

[0013] "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and must be harmless to the recipient.

[0014] The present disclosure also relates to a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier or diluent or an effective amount of a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier or diluent.

[0015] These embodiments generally refer to a new process for the preparation of tin mesoporphyrin halides. In embodiments, the tin mesoporphyrin compound may be formed by heme transmetalation followed by hydrogenation of tin protoporphyrin to provide a tin mesoporphyrin halide. In other embodiments, a method of synthesizing a tin mesoporphyrin halide comprises forming the protoporphyrin methyl ester from hemin and converting the medium-protoporphyrin methyl ester product to the mesoporphyrin tin halide by introducing tin and hydrogenation.

[0016] IX tin tinoprofirin dichloride or stannsoporfin is a chemical compound with the structure shown in FIG. 3. It is recommended to use it, for example, as a medicine for the treatment of various diseases including, for example, psoriasis and jaundice. Stannsoporfin may also lead to inhibition of heme metabolism in mammals, to control the rate of tryptophan metabolism in mammals, and to increase the rate at which heme is excreted by mammals.

The incorporation of metal into mesoporphyrin IX dihydrochloride to yield a mesoporphyrin metal halide is described with particular reference to tin to provide stannsoporfin, a known drug and a particularly preferred embodiment of the invention.

[0018] In embodiments, the tin mesoporphyrin compound may be formed by heme transmetalation followed by hydrogenation of tin protoporphyrin to provide a tin mesoporphyrin halide. In embodiments, the heme is transmetalised using tin to provide tin protoporphyrin and then hydrogenated in a solvent such as n-methylpyrrolidone, dilute ammonium hydroxide, or dimethylformamide. In some embodiments, the hemin can be trans metallated with or without iron sulfate. In some embodiments, the hemin subjected to transmetalation is treated with charcoal. In some embodiments, the product is isolated after hydrogenation by the addition of acetic acid or hydrochloric acid. In some embodiments, the final product is precipitated using MTBE. In some embodiments, the product is further purified using chromatography.

[0019] In other embodiments, a method of synthesizing a tin mesoporphyrin halide comprises forming the protoporphyrin methyl ester from hemin and converting the intermediate methyl ester of protoporphyrin to the mesoporphyrin halide by introducing tin and hydrogenation. To obtain the methyl ester, hemin, pyridine and dichloromethane, it can be stirred until a solution is obtained. Iron sulphate, methanol, dichloromethane and HCl gas can then be added to give an exothermic reaction. In exemplary embodiments, the exothermic reaction may then be maintained at reflux for a period of about 2 to about 5 hours. In the embodiments, the reaction may then be washed with water and diluted using ammonium hydroxide to provide the protoporphyrine methyl ester. In some embodiments, the tin may be incorporated into the protoporphyrine methyl ester using tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2-ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate. In embodiments, the tin protoporphyrin dimethyl ester can be hydrogenated using a suitable metal catalyst under a hydrogen atmosphere to provide dimethyl mesoporphyrin tin ester. In some embodiments, the tin protoporphyrin dimethyl ester may be hydrogenated to provide a tin mesoporphyrin dimethyl ester. In some embodiments, the tin mesoporphyrin halide can be obtained by heating tin mesoporphyrin dimethyl ester in dilute ammonium hydroxide. In some embodiments, the stannsoporfin can be obtained from tinoporphyrin dimethyl ester by heating the material to from about 70 ° C to about 80 ° C in dilute ammonium hydroxide. For example, stannsoporfin can be obtained by heating dimethyl tin mesoporphyrin to about 75 ° C in dilute ammonium hydroxide. In some embodiments, the product may be further purified using chromatography.

[0020] Also described is a method for synthesizing a metal mesoporphyrin halide comprising forming an intermediate mesoporphyrin IX dihydrochloride and converting an intermediate mesoporphyrin IX dihydrochloride to a mesoporphyrin metal halide by metal insertion. The hemine can be hydrogenated in formic acid with a suitable metal catalyst under a hydrogen atmosphere from about 80 ° C to about 101 ° C for about 1 hour to about 3 hours. The hydrogenation of the hemin can be continued for an additional time of about 24 hours to about 36 hours at about 40 ° C to about 60 ° C. For example, hemin can be hydrogenated at about 85 ° C to about 90 ° C at about 60 psi of hydrogen for about 1 hour to about 2 hours and then at about 45 ° C to about 50 ° C for about 24 hours to about 36 hours. The metal catalyst may be palladium, nickel, platinum, palladium on carbon or the like. After the hydrogenation has been completed, the reaction may be cooled to about 20 ° C to about 30 ° C or about 20 ° C to about 25 ° C and may optionally be supplemented with powdered activated carbon, such as that sold under the trade name Darco KB-G. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, e.g., by vacuum distillation. A solution of about 1N HCl can then be added over one hour or more to precipitate the intermediate. After filtration, the product is dried under a stream of nitrogen to obtain mesoporphyrin IX dihydrochloride. After the hydrogenation has been completed, the reaction may be cooled to about 20 ° C to about 30 ° C or about 20 ° C to about 25 ° C and may optionally be supplemented with powdered activated carbon, such as that sold under the trade name Darco KB-G. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, e.g., by vacuum distillation. A solution of about 1N HCl can then be added over one hour or more to precipitate the intermediate. After filtration, the product is dried under a stream of nitrogen to obtain mesoporphyrin IX dihydrochloride. After the hydrogenation has been completed, the reaction may be cooled to about 20 ° C to about 30 ° C or about 20 ° C to about 25 ° C and may optionally be supplemented with powdered activated carbon, such as that sold under the trade name Darco KB-G. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, e.g., by vacuum distillation. A solution of about 1N HCl can then be added over one hour or more to precipitate the intermediate. After filtration, the product is dried under a stream of nitrogen to obtain mesoporphyrin IX dihydrochloride. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, e.g., by vacuum distillation. A solution of about 1N HCl can then be added over one hour or more to precipitate the intermediate. After filtration, the product is dried under a stream of nitrogen to obtain mesoporphyrin IX dihydrochloride. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, e.g., by vacuum distillation. A solution of about 1N HCl can then be added over one hour or more to precipitate the intermediate. After filtration, the product is dried under a stream of nitrogen to obtain mesoporphyrin IX dihydrochloride.

[0021] A second method step according to one or more embodiments of the invention is shown in FIG. 1 as RPA438-03-ES with reference to tin as the metal introduced (standard tin oxide path). The metal may include tin, iron, zinc, chromium, manganese, copper, nickel, magnesium, cobalt, platinum, gold, silver, arsenic, antimony, cadmium, gallium, germanium, palladium and the like. The metal carrier may be a tin (II) carrier. Carriers such as tin (II) halide or tin (II) acetate may be used. The tin carrier may be tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2-ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate. In the standard tin oxide path, tin (II) oxide powder can be suspended in acetic acid. A solution of mesoporphyrin IX dihydrochloride in formic acid under nitrogen can be added to this suspension and heated to from about 60 ° C to about 65 ° C. After completion of the reaction, water can be added slowly, and the reaction can be slowly cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered. The filtering mass may be washed with water, suspended in 1 N HCl and heated to from about 85 ° C to about 95 ° C for from about 1 hour to about 3 hours. The slurry may be cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered and dried under nitrogen to give the crude tin di mesoporphyrin IX dihydrochloride. and the reaction can be slowly cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered. The filtering mass may be washed with water, suspended in 1 N HCl and heated to from about 85 ° C to about 95 ° C for from about 1 hour to about 3 hours. The slurry may be cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered and dried under nitrogen to give the crude tin di mesoporphyrin IX dihydrochloride. and the reaction can be slowly cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered. The filtering mass may be washed with water, suspended in 1 N HCl and heated to from about 85 ° C to about 95 ° C for from about 1 hour to about 3 hours. The slurry may be cooled to from about 20 ° C to about 30 ° C or from about 20 ° C to about 25 ° C, stirred and filtered and dried under nitrogen to give the crude tin di mesoporphyrin IX dihydrochloride.

[0022] For example, powdertin (II) oxide (SnO, 1.7 kg) can be suspended in about 40.5 L of acetic acid to from about 20 ° C to about 25 ° C and then heated to from about 60 ° C to about 65 ° C in an atmosphere. nitrogen. To this suspension, a solution of mesoporphyrin IX dihydrochloride (B991, 2.1 kg) in about 10.5 L of formic acid can be added over a period of about 6 hours. The reaction may be carried out under a nitrogen atmosphere at from about 60 ° C to about 65 ° C for at least about 12 hours. The reaction can be monitored using HPLC. After the reaction, about 17 l of water can be added over 0.5 hour. The reaction mixture is then cooled to from about 20 ° C to about 25 ° C for about 0.5 hour, stirred for about 1 to about 3 hours and then filtered. The filtering mass can then be washed with distilled water (USP), suspend in about 1N HCl and heat to from about 85 ° C to about 95 ° C for about 1 to about 3 hours. The slurry may then be cooled to from about 20 ° C to about 25 ° C, stirred for about 0.5 hour, filtered, rinsed with distilled water (USP) and dried under nitrogen to give about 1.3 to about 1.5 kg of crude IX tin mesoporphyrin dichloride (stannsoporfin).

[0023] The tin mesoporphyrin dihydrochloride can be heated with a metal carrier in acetic acid in the presence of an oxidation at the reflux (oxidation process with reflux). The metal may include tin, iron, zinc, chromium, manganese, copper, nickel, magnesium, cobalt, platinum, gold, silver, arsenic, antimony, cadmium, gallium, germanium, palladium and the like. The metal carrier may be a tin (II) carrier. Carriers such as tin (II) halides or tin (II) acetate may be used. The tin carrier may be tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate.

[0024] Heating is carried out, for example, with air purging, for example by inflow of about 6% oxygen mixed with nitrogen for about 24 hours to about 48 hours. The air supply can also be used for air blowing during heating. The reaction can also be carried out in the presence of suitable acetate counterions, including ammonium, sodium or potassium ions. It is also possible to use oxygen from the air in both pure form and as hydrogen peroxide. For example, mesoporphyrin IX formate is heated using tin (II) oxide in acetic acid, buffered with ammonium acetate, and the reaction is carried out using aeration and backflow (reflux). Ammonium acetate can be eliminated. Metallized methylporphyrin can be isolated from the reaction mixture by adding water, and then filtration. Prior to drying, the mass may be rubbed in hot dilute hydrochloric acid, preferably at a concentration of about 0.1N 6N and at a raised temperature of 90 ° C to 100 ° C. The reaction may give the metal mesoporphyrin IX crude dichloride. The reaction may give raw tin IX mesoporphyrin dichloride.

The metal mesoporphyrin dichloride obtained in this way can then be purified by dissolving the product in an aqueous solution of an inorganic base, preferably dilute ammonium hydroxide, followed by subsequent treatment with charcoal. The product can then be re-precipitated by introducing into the solution an acid such as acetic acid, hydrochloric acid or mixtures thereof. The above solubilizing treatment using charcoal treatment and re-precipitation steps can be repeated many times, usually about 1 to 3 times to ensure the desired purity. Prior to drying, the mass is triturated in hot dilute hydrochloric acid at a concentration of about 0.1 N to about 6 N at an elevated temperature of about 90 ° C to about 100 ° C to remove any residual ammonium salts. A mesoporphyrin chloride methylation product (tin IX mesoporphyrin tin or stannsoporfin dichloride) can be obtained. A tin product of mesoporphyrin chloride (tin mesoporphyrin IX tin or stannsoporfin dichloride) can be obtained. The end product, e.g. stannsoporfin, can be isolated using chromatography.

[0026] For example, tin mesoporphyrin tin dichloride (1.7 kg) in 2% ammonium hydroxide (22 L) can be dissolved using tin mesoporphyrin dichloride. A pH measurement can be made to ensure that the pH is> 9.0. The solution can be processed using Darco KB-G (0.1 kg) and Hyflo Supercel (0.2 kg), stirred for 1 to 2 hours and filtered to remove solids. The filtrate may then be added dropwise to acetic acid (44 L) containing hydrochloric acid (31%, 2.7 L) while maintaining the temperature at 20 ° C to about 25 ° C. The pH measurement can be repeated again to ensure that the pH is <1.0. The resulting suspension can be stirred for 1 to 2 hours under nitrogen to isolate the product by filtration. The wet mass can then be triturated in 3N HCl (35 L) at 85 ° C to about 90 ° C and stirred for about 16 hours to about 18 hours to convert the crystalline form to the monomer and remove the residual ammonium salts. The suspension may be cooled to from 20 ° C to about 25 ° C and the product isolated by filtration. The mass of the product may be washed with 0.3N HCl (16 L) and dried under a stream of nitrogen to obtain from about 1.2 to about 1.6 kg of Stannsoporfin.

[0027] The metal may include tin, iron, zinc, chromium, manganese, copper, nickel, magnesium, cobalt, platinum, gold, silver, arsenic, antimony, cadmium, gallium, germanium, palladium and the like. The preparation of the mesoporphyrin halides of these other metals includes simply substituting a halide such as chloride, bromide or iodide of the chosen metal in place of tin (II) chloride in the manner described, in substantially equivalent amounts.

The metal mesoporphyrin halide can be formed from hemin by anhydrogen-hydrogenation process to give the intermediate mesoporphyrin IX, then metal is introduced into the mesoporphyrin IX intermediate and the metallized intermediate is hydrogenated to give the metal mesoporphyrin halide. Mesoporphyrin can be obtained by reacting hemin with iron sulphate, palladium on carbon and poly (methylhydrosiloxane) (PMHS) in formic acid with reflux and subsequent hydrogenation using PMHS. The product can be isolated as formate of mesoporphyrin from formic acid and methyl tert-butyl ether. The product can then be passed through the above-described process to obtain the dihydrochloride. The hemoline can be reacted with iron sulphate, palladium on carbon and poly (methylhydrosiloxane) (PMHS) in formic acid and then concentrated using a rotary evaporator to remove formic acid. The solids obtained in this way can be isolated by filtration and through a standard tin oxidation pathway. The filtrate obtained may then be concentrated and dissolved in dilute ammonium hydroxide and precipitated by the addition of acetic acid or hydrochloric acid. The iron can be removed from the hemin with iron sulphate before PMHS hydrogenation. The product can then be purified using chromatography. The filtrate obtained may then be concentrated and dissolved in dilute ammonium hydroxide and precipitated by the addition of acetic acid or hydrochloric acid. The iron can be removed from the hemin with iron sulphate before PMHS hydrogenation. The product can then be purified using chromatography. The filtrate obtained may then be concentrated and dissolved in dilute ammonium hydroxide and precipitated by the addition of acetic acid or hydrochloric acid. The iron can be removed from the hemin with iron sulphate before PMHS hydrogenation. The product can then be purified using chromatography.

[0029] In any of the above embodiments, the reagents, intermediates and / or products may pass through additional purification steps. In some embodiments, the additional purification involves treating the reagent, intermediate, or product using diatomaceous earth and / or activated carbon. In one embodiment, treating the reagent, intermediate product, or product with diatomaceous earth and / or activated carbon involves dissolving or suspending the reagent, intermediate product and / or product in a solvent, adding diatomaceous earth and / or activated charcoal, filtering off kieselguhr and / or carbon activated and recovering the reagent, intermediate or product from the filtrate. In some embodiments, the additional purification involves triturating the reagent, intermediate or product in a hot acid such as about 0.1 to 6N HCl in water, preferably about 3N HCl in water. In some embodiments, one, two or three diatomaceous earth treatment steps, activated carbon treatments and hot acid triturations are carried out one after another in any order and can be repeated if desired.

Metal mesoporphyrin halide can be obtained directly from hemin without isolation of any intermediates. The mesoprofirin compound of the metal can be synthesized without isolating the mesoporphyrin formate intermediate or the mesoporphyrin dihydrochloride intermediate. Metal mesoporphyrin can be synthesized using any of the methods described above without isolating the intermediate mesoporphyrin dihydrochloride product. Metal mesoporphyrin can be synthesized using any of the methods described above without isolating the intermediate. Metal mesoporphyrin can be synthesized using the standard tin oxide route or the oxidation method with reflux as described above without isolating the intermediate product of the dihydrochloride dihydrochloride. Metal mesoporphyrin can be synthesized using the standard tin oxide route or the oxidation method with reflux as described above without isolating the intermediate. The method for synthesizing stannsoporfin may include hemin hydrogenation and heating the reaction in the presence of a metal carrier. Heating takes place under a nitrogen atmosphere. The method for synthesizing stannsoporfin may include hydrogenation of hemin and heating the resulting reaction using a metal carrier and acetic acid in the presence of an oxidizing agent with reflux. The metal may include tin, iron, zinc, chromium, manganese, copper, nickel, magnesium, cobalt, platinum, gold, silver, arsenic, antimony, cadmium, gallium, germanium, palladium and the like. The metal carrier may be a tin (II) carrier. Carriers such as tin (II) halide or tin (II) acetate may be used. The tin carrier may be tin oxide, tin chloride, tin sulfate,

[0031] Hemin may be hydrogenated in formic acid with a suitable metal catalyst under a hydrogen atmosphere from about 80 ° C to about 101 ° C for about 1 hour to about 3 hours. The hydrogenation of the hemin can be continued for an additional time of about 24 hours to about 36 hours at about 40 ° C to about 60 ° C. For example, hemin can be hydrogenated at about 85 ° C to about 90 ° C at about 60 psi of hydrogen for about 1 hour to about 2 hours and then at about 45 ° C to about 50 ° C for about 24 hours to about 36 hours. The metal catalyst may be palladium, nickel, platinum, palladium on carbon or the like. After completion of the hydrogenation, the reaction may be cooled to about 20 ° C to about 30 ° C or about 20 ° C to about 25 ° C and optionally it may be supplemented with powdered activated carbon, such as that sold under the trade name Darco KB-G. The reaction may optionally be stirred prior to filtration. The reaction can be filtered through a metal eliminating agent such as Hyflo Supercel to remove the catalyst. The filtered solution can optionally be concentrated, for example, by vacuum distillation.

[0032] Reagents, intermediates and / or products may pass through additional purification steps. Additional purification may include treatment of the reagent, intermediate product or product with diatomaceous earth and / or activated charcoal. Treatment of the reagent, intermediate product or product with diatomaceous earth and / or activated carbon may involve dissolving or suspending reagent, intermediate and / or product in a solvent, adding diatomaceous earth and / or activated charcoal, filtering off kieselguhr and / or activated carbon and recovering the reagent , an intermediate product or a product from the filtrate. Additional purification may include triturating the reagent, intermediate or product in a hot acid such as from about 0.1 to about 6N HCl in water, preferably about 3N HCl in water. One,

[0033] A method of obtaining a metal mesoporphyrin halide has been described which comprises the steps of: a) exposing a hydrogen hydrogen hydrogen metal hydrogenation catalyst to obtain a prehydrogenated catalyst; and b) joining the hemin with a pre-hydrogenated catalyst and maintaining the hemin and catalyst in one or more cycles at a combination of temperature, hydrogen pressure and time that are sufficient to remove the iron from the heme and reduce the hemin vinyl groups to the ethyl groups, thereby obtaining mesoporphyrin IX. Also described is a method for obtaining a metal mesoporphyrin halide, which comprises the steps of: a) exposing the hydrogen hydrogen hydrogen metal catalyst to obtain a pre-hydrogenated catalyst;

Step b) can be performed at about 80 ° C to about 100 ° C, preferably at about 85 ° C to about 90 ° C, at a hydrogen pressure from about 50 to about 70 psi, preferably from about 55 to about 60 psi, from about 1 to about 3 hours, preferably about 1 to about 1.5 hours; then at about 40 ° C to about 60 ° C, preferably at about 45 ° C to about 50 ° C, at a hydrogen pressure from about 50 to about 70 psi, preferably from about 55 to about 60 psi, for about 18 to about 48 hours, preferably about 24 hours.

[0035] The metal hydrogenation catalyst may include palladium, palladium on carbon, platinum, platinum on carbon, nickel or nickel aluminum catalyst. The metal hydrogenation catalyst may be palladium. The metal hydrogenation catalyst may be palladium on carbon.

[0036] The methods described herein also include the introduction of tin into mesoporphyrin to provide stannsoporfin using tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2-ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate. The tin can be incorporated into the mesoporphyrin using tin oxide, tin chloride or tin 2-ethylhexanoate.

[0037] Mesoporphyrin IX hydrochloride can be treated using a tin (II) salt in an organic solvent such as acetic acid under oxidative conditions that yield the desired product, tin (IV) mesoporphyrin IX mesodhorin (stannsoporfin). For example, mesoporphine IX dihydrochloride and tin (II) chloride may be placed in a reservoir and acetic acid may be added at about 20 ° C to about 30 ° C, preferably at about 20 ° C to about 25 ° C. The suspended reagents are mixed for at least about 30 minutes. During vigorous stirring, the mixture is heated in inert fumes (such as nitrogen or argon) at the reflux.

[0038] Also described is a method for introducing tin into mesoporphyrin IX comprising reacting mesoporphyrin IX with tin salt in the presence of a proton elimination agent.

[0039] Also described is a method for introducing tin into mesoporphyrin IX, comprising reacting mesoporphyrin IX with tin salt at the rate of oxidation. Mesoporphyrin IX can be reacted with tin salt in the reaction vessel with the free space at the top, and the rate of oxidation is controlled by introducing a gas containing oxygen into the free space of the reaction vessel. The oxygen-containing gas introduced into the free space of the reaction vessel can have about 3% to about 22% oxygen in an inert gas such as nitrogen. The oxygen-containing gas introduced into the free space of the reaction vessel can be air. The oxygen-containing gas introduced into the free space of the reaction vessel may have about 4% to about 15% oxygen in an inert gas such as nitrogen. The oxygen-containing gas introduced into the free space of the reaction vessel can have about 5% to about 10% oxygen in an inert gas such as nitrogen. The oxygen-containing gas introduced into the free space of the reaction vessel may have about 6% oxygen in an inert gas such as nitrogen. The oxygen-containing gas introduced into the free space of the reaction vessel may have about 6% oxygen in nitrogen.

[0040] The methods described herein also include the introduction of tin into mesoporphyrin to provide stannsoporfin using tin oxide, tin chloride, tin sulfate, tin bromide, tin oxalate, tin pyrophosphate hydrate, tin 2-ethylhexanoate, methanesulfonic acid tin, or tin trifluoromethanesulfonate.

[0041] Other porphyrin compounds and tetrapyrroles can also be metered using the procedures described herein including, but not limited to, porphyrins such as deuteroporphyrins and deuteroporphyrin IX 2,4-bis (ethylene glycol) (8,13-bis (1,2-dihydroxyethyl) 3 acid). , 7,12,17-tetramethyl-21H, 23H-porphine-2,18-dipropionowego). Additional porphyrin compounds that can be metered using the procedures described herein include, but are not limited to, coproporphyrin, cytoporphyrin, ethioporphyrin, hematoporphyrin, mesoporphyrin, filoporphyrin, protoporphyrin, piroporifyrine, rhodoporphyrin, uroporphyrin and phytoporphyrin. A comprehensive list of porphyrin compounds can be found on the website, chem.qmul.ac.uk/iupac/tetrapyrrole/; the porphyrins described therein are incorporated herein by reference as porphyrins that can be metered using the procedures described herein.

[0042] This invention and the embodi- ments used to illustrate the method and materials used can be further understood by reference to the following non-limiting examples.

EXAMPLE 1 [0043] Stannsoporfins can be obtained via the inverse pathway, where the heme is trans metallated using tin oxide to form tin protoporphyrin, followed by hydrogenation in n-methylpyrrolidone, dilute ammonium hydroxide or dimethylformamide.

[0044] A reaction was carried out whereby the hemin was reacted with the tin oxide RPA438-03-ES (see FIG 1) with and without the addition of ferric sulphate. After heating the reaction mixture at 90 ° C for 2 hours under nitrogen to effect dissolution of hemin, the reaction was continued at 60 ° C to about 65 ° C overnight. HPLC analysis confirmed the completion of the reaction for the reaction without iron sulfate and completion in 98.4% for the reaction with iron sulphate. Both reactions were isolated by standard addition of water at an identical yield of 97.3%.

[0045] Based on the success of these experiments, the reaction without iron sulphate was scaled to 40 g to obtain material for testing subsequent transformations. For the results, refer to Table 1.

<td colspan="6">TABLE 1: ANALYTICAL RESULTS, PROTOPORPHYHINE</td>

<td>Experience</td><td>Description</td><td>Whole Yield (%) for Hemina</td><td>Cleanliness with HPLC (% a / a)</td><td>tested HPLC w / w)</td><td>quantity (%</td>

<td>1165-CB-1411</td><td>protoporphyrin tin</td><td>92.5</td><td>98.1</td><td>104.9</td><td></td>

[0046] THICKNESSED AMMONIUM HYDROXIDE: Hydrogenation was carried out in dilute ammonium hydroxide on a palladium catalyst. The solvent volume and concentration were chosen to be appropriate for the RPA438-04-EF purification process depicted in FIG. 1.

Initial IPC analysis for this reaction showed no reaction. However, after isolating and analyzing the product using a method for determining the final purity of the product, it was determined that the solid contained 18.2% stannsoporfin (B992; FIG. 3), 14.7 and 39.5% monovinyl intermediates (CJ9 / CKO; 4) and 27.6% tin protoporphyrin (CH8; 5).

[0048] The reaction of ammonium hydroxide was repeated using pre-treatment using charcoal. The first IPC analysis after overnight reaction showed 83.3% stannsoporfin (B992), 15.9% monovinyl intermediates (CJ9 / CKO) and 0.7% tin protoporphyrin (CH8). The reaction mixture was filtered and the hydrogenation continued with fresh catalyst to complete the reaction in less than 2 hours. The products were isolated by introducing into acetic acid / hydrochloric acid (as in the RPA438-04-EF purification process depicted in FIG 1) and triturated in hot 1N HCl for 1 hour. The efficiency of this transformation was 78%. The product from this reaction was passed through a purification process with a yield of 91.4%. The total process yield was 65.9% with a purity of 98.8% (See Table 2, Ref .: 1165-CB-172-1).

[0049] DIMETHYLFORMAMID: Hydrogenation was performed using a dimethylformamide solvent (DMF) and palladium on carbon. Both the starting material as well as the product can be dissolved in 50 parts, allowing the catalyst to be removed after completion of the reaction. After overnight reaction during the analysis of the process (IPC), the analysis showed 35.5% B992, 28 and 11% monovinyl intermediates and 25.4% tin protoporphyrin. The DMF solution in the IPC step was analyzed by mass spectrometry with liquid chromatography (LC / MS) to confirm the identity of each peak with IPC HPLC.

[0050] N-METYLOPIROLIDON (NMP): NMP was defined as the hydrogenation solvent. After overnight reaction at 50 ° C / 60 psi, the IPC analysis showed 51% stannsoporfin (B992), 20% and 10% monovinyl intermediates (CJ9 / CKO) and 18.5% tin protoporphyrin (CH8). During this reaction, it was observed that all hydrogen was collected during the first 1 to 2 hours of reaction, which indicates that the catalyst may have been poisoned. The reaction was then reheated to 90 ° C / 60 psi overnight. IPC analysis showed that the reaction continued to 68% stannsoporfin (B992), 21 and 9% monovinyl intermediates (CJ9 / CKO) and 2.1% tin protoporphyrin (CH8), which indicates that the catalyst did not completely poison .

[0051] Assuming the catalyst poisoned during the reaction, the NMP protoporphyrin tin solution (CH8) was treated with Darco-KBG activated charcoal for 1 hour and filtered before introducing the hydrogenation catalyst. After an overnight reaction at 50 ° C / 60 psi, the IPC analysis showed 94.2% stannsoporfin (B992) and 1.7% monovinyl intermediates (CJ9 / CKO). The reaction mixture was divided into two equal parts and added to 300 parts of water or 300 parts of methyl tert-butyl ether (MTBE) and cooled to from about 0 ° C to about 5 ° C for about 2 hours. After filtration, no precipitate was found in the water or was low, but MTBE gave a well-filtered solid at a 90% yield.

[0052] Hydrogenation in NMP with charcoal treatment was increased to 10 g. The first IPC analysis after reaction at 50 ° C / 60 psi overnight showed 92.5% stannsoporfin (B992), 3.6 and 2.4% monovinylated products intermediate (CJ9 / CKO) and 0.4% tin protoporphyrin (CH8). The reaction mixture was filtered before re-hydrogenation using fresh catalyst, bringing the reaction to completion within the next 2 hours. The product was isolated by adding abundant amounts of methyl tert-butyl ether at a yield of 119.9% ​​(the product contains residual NMP). The product from this reaction was passed through a purification process at a yield of 108.0% (probably due to residual solvents). The purity obtained was 98.2%. (See Table 2, Ref .: 1165-CB-171-1).

<td colspan="8">TABLE 2: ANALYTICAL RESULTS, REVERSE PATH</td>

<td>Experience</td><td>Description</td><td>Total Efficacy (%) for Hemina</td><td colspan="4">HPLC purity (% A / a)</td><td>tested HPLC amount (% W / w)</td>

<td>1165-CB-1711</td><td>stannsoporfin Obtaining the reverse path Hydrogenation of the Virgin Mary</td><td>119.9</td><td colspan="4" rowspan="2">98.2</td><td>47.0</td>

<td></td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>RRT</td><td>% A / a</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.06</td><td>0.13</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.25</td><td>0.11</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.42</td><td colspan="2">0.20</td><td></td>

<td></td><td></td><td></td><td></td><td>0.62</td><td>0.23</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.66</td><td>0.10</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.88</td><td>0.10</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.9</td><td>0.10</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>Other</td><td>&lt;0.10</td><td></td><td></td>

<td>1165-CB-1721</td><td>Stannsoporfin. Obtaining the inverse path The hydrogenation NH4OH</td><td>65.9</td><td colspan="4">98.8</td><td>80.8</td>

<td></td><td></td><td></td><td></td><td>RRT</td><td>% A / a</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.09</td><td>0.13</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.25</td><td>0.09</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.88</td><td>0.10</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.9</td><td>0.05</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>Other</td><td>&lt;0.14</td><td></td><td></td>

EXAMPLE 2 [0053] Obtaining Methyl Protoporphyrin Esters. To the 5 L reactor, 20 g of hemin, 50 ml of pyridine and 200 ml of dichloromethane were added and mixed for 10 minutes to obtain a solution. Then 50 g iron sulfate, 1000 ml methanol and 1000 ml dichloromethane were added. HCl gas was added slowly. The exothermic reaction finally reached a reflux temperature of 41 ° C (note: no heating was given) and kept at reflux for 3 hours. HPLC analysis confirmed the completion of the reaction. The reaction was quenched with 1000 ml of water, followed by additional washing with water and dilute ammonium hydroxide. The protoporphyrin methyl ester was then isolated from dichloromethane / methanol at a yield of 50.4%.

[0054] Introduction of tin with subsequent hydrogenation. Dimethyl protoporphyrin tin ester was obtained according to the standard tin oxide pathway directly substituting the protoporphyrin dimethyl ester as mesoporphyrin. The product was obtained in a yield of 77.7%. [0055] Tin protoporphyrin dimethyl ester was hydrogenated in dichloromethane in a 5% palladium catalyst at 50 psi. After the reaction overnight, the product was isolated by concentration using a rotary evaporator at 100% yield. NMR analysis of the product indicates that there is about 10% unreacted starting material. However, after re-exposure of the material to the hydrogenation conditions, no further changes were observed and the reaction was continued under such conditions.

[0056] Stannsoporfin was obtained from mesoporphyrin dimethyl ester by heating the material to 75 ° C in dilute ammonium hydroxide. After 18 hours of reaction, no further changes in IPC HPLC were observed even after the addition of further amounts of ammonium hydroxide. The product was isolated from the solution by addition to acetic acid / hydrochloric acid, which was usually refined during purification, with a yield of 72.6%. The crude Stannsoporfin was purified according to a standard method and isolated at a yield of 72.2%. The total process yield was 23.8% with a purity of 87.4% (See Table 3, Ref .: 1198-CB-003-1).

TABLE 3: RESULTS OF ANALYSIS, PATH OF METALLY ETHER

<td>Experience</td><td>Description</td><td>Total Efficacy (%) for Hemina</td><td colspan="4">HPLC purity (% A / a)</td><td>The quantity tested HPLC (current % W / w)</td>

<td></td><td></td><td></td><td colspan="4">87.4</td><td></td>

<td></td><td></td><td></td><td></td><td>RRT</td><td>% A / a</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.27</td><td>0.18</td><td></td><td></td>

<td rowspan="2">1198-CB-0031</td><td>stannsoporfin</td><td></td><td></td><td>0.42</td><td>0.24</td><td></td><td></td>

<td rowspan="3">receiving methyl ester</td><td rowspan="2"></td><td rowspan="2"></td><td rowspan="2">0.96</td><td rowspan="2">2.06</td><td rowspan="2"></td><td rowspan="2"></td>

<td></td>

<td></td><td></td><td></td><td>1.78</td><td>9.90</td><td></td><td></td>

<td></td><td></td><td>23.8</td><td></td><td>Other</td><td>&lt;0.05</td><td></td><td>82.8</td>

EXAMPLE 3 [0057] Mesoporphyrin was obtained by reacting hemin with iron sulphate, palladium on carbon and poly (methylhydrosiloxane) (PMHS) in formic acid with reflux. The reaction was separated. Half was further hydrogenated using PMHS. The IPC then showed the reaction was complete. The product was isolated as CK1 (structure shown in FIG 1) from formic acid and tert-butyl methyl ether (1198-CB004-1). The insulation was very difficult due to solids (both products and waste) that adhered to the walls of the flask. This is probably due to the side product formed from PMHS, which is considered a silicone paste. The initial reaction gave 57.5% of mesoporphyrin.

[0058] In the second half of the reaction, attempts were made to directly isolate the B991 hydrochloride salt. However, no crystals were observed after the standard method. The solution was concentrated using a vacuum evaporator to remove formic acid, and the remaining solids were isolated by filtration at a yield of 42.8%. The second batch of material was obtained from the filtrate the next day with an additional yield of 11.2%. Two batches of material were combined and introduced into the tin oxide process. After the standard reaction time no further B991 was detected using HPLC. Attempts have been made to isolate the product according to a standard process, however, no solids have been detected. The filtrate was then concentrated using a vacuum evaporator, and the precipitate was dissolved in dilute ammonium hydroxide and precipitated by addition of acetic acid / hydrochloric acid. The crude product was obtained at a yield of 26.6% (Table 4, Ref .: 1198-CB-011-1).

[0059] Attempts have also been made to remove iron from hemin using iron sulphate before hydrogenation of PMHS. The reaction is easier to carry out because iron salts and soaps, both insoluble, did not occur in the reactor at the same time. The product was isolated with a yield of 62.1%. However, when attempts were made to obtain B991 (the structure shown in FIG 1) on this material, it was noted that no solids were precipitated after the addition of hydrochloric acid to the formic acid solution.

The product from reaction 1198-CB-004-1 was performed by the formation of B991 hydrochloride in a yield of 42.5%. The tin oxide method was repeated with this sample, but the precipitate was stirred overnight before filtering. The product from this reaction was obtained at a yield of 90.3%. The total yield was 22.1% at 75.1% purity. (Table 4, Ref .: 1198-CB-013-1).

<td colspan="8">TABLE 4: RESULTS OF ANALYSIS, RECEIVING NO HYDROGEN</td>

<td>Experience</td><td>Description</td><td>Total Efficacy (%) for Hemina</td><td colspan="4">HPLC purity (% A / a)</td><td>The quantity tested HPLC (current % W / w)</td>

<td>1198-CB-0111</td><td>stannsoporfin</td><td>14.4</td><td colspan="4">73.3</td><td>25.2%</td>

<td></td><td>receiving bezwodorowe</td><td></td><td></td><td>RRT</td><td>% A / a</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.38</td><td>0.63</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.42</td><td>0.32</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.70</td><td>0.44</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.76</td><td>0.41</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.79</td><td>0.54</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>0.89</td><td>21.95</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.71</td><td>0.31</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.14</td><td>0.26</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>Other</td><td>&lt; 0.30</td><td></td><td></td>

<td>1198-CB-0131</td><td>stannsoporfin</td><td>22.1</td><td colspan="4">75.1</td><td>0.8%</td>

<td></td><td>receiving bezwodorowe</td><td></td><td></td><td>RRT</td><td>% A / a</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.61</td><td>2.36</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.70</td><td>3.39</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.74</td><td>0.71</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.78</td><td>3.63</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.80</td><td>2.64</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.87</td><td>0.63</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.92</td><td>2.57</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>1.96</td><td>0.89</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.02</td><td>0.33</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.06</td><td>1.95</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.15</td><td>1.65</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.17</td><td>1.37</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.19</td><td>0.44</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.21</td><td>0.80</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>2.30</td><td>1.35</td><td></td><td></td>

<td></td><td></td><td></td><td></td><td>Other</td><td>&lt; 0.20</td><td></td><td></td>

EXAMPLE 4 [0061] Each tin carrier was evaluated for reflux oxidation for obtaining stannsoporfin shown in FIG. 2 and the low-temperature anaerobic method described in FIG. 1. Initial selection reactions were performed on a 150 mg scale. The results of the initial selection reactions regarding the oxidation process and the low-temperature anaerobic process are summarized in Tables 5 and 6, respectively

<td colspan="6">TABLE 5: IPC RESULTS, ALTERNATIVE SALTS IN THE OXIDIZATION METHOD</td>

<td rowspan="2">Experience</td><td rowspan="2">Tested tin salts</td><td colspan="2">24 hours</td><td colspan="2">48 hours</td>

<td>B991 terms of</td><td>The peak cleanliness</td><td>B991 terms of</td><td>The peak cleanliness</td>

<td></td><td></td><td>B992 (%)</td><td>B992 (% A / a)</td><td>B992 (%)</td><td>B992 (% A / a)</td>

<td>1198-CB-015</td><td>Tin sulphate (II)</td><td>215.95</td><td>25.19</td><td>163.6</td><td>36.06</td>

<td>1198-CB-086-1</td><td>Tin oxide (II)</td><td>0.11</td><td>99.48</td><td>0.22</td><td>98.63</td>

<td>1198-CB-086-2</td><td>Tin bromide (II)</td><td>13.65</td><td>84.74</td><td>12.29</td><td>85.99</td>

<td>1198-CB-086-3</td><td>Tin oxalate (II)</td><td>49.5</td><td>62.76</td><td>48.6</td><td>61.05</td>

<td>1198-CB-086-4</td><td>Tin sulphate (II)</td><td>Lack reaction</td><td>0.00</td><td>10000,00</td><td>0.96</td>

<td>1198-CB-086-5</td><td>Pyrophosphate hydrate tin (II)</td><td>79.26</td><td>54,44</td><td>34,09</td><td>73.3</td>

<td>1198-CB-086-6</td><td>Tin (II) ethylhexanoate</td><td>0.04</td><td>99.21</td><td>0.16</td><td>99.09</td>

<td>1198-CB-086-7</td><td>Methanesulfonic acid tin (II) (50% solution in water)</td><td>627</td><td>13.72</td><td>337.5</td><td>12.43</td>

<td>1198-CB-086-8</td><td>Tin trifluoromethanesulfonate (II)</td><td>Lack reaction</td><td>0.00</td><td>1,007.95</td><td>6.16</td>

<td colspan="6">TABLE 6: IPC RESULTS, ALTERNATIVE SALT IN THE LOW-TEMPERATURE METHOD</td>

<td>experienced</td><td>Tested tin salts</td><td colspan="2">24 hours</td><td colspan="2">48 hours</td>

<td>e</td><td></td><td>B991</td><td>summits</td><td>B991</td><td>summits</td>

<td></td><td></td><td>terms of</td><td>and cleanliness</td><td>terms of</td><td>and cleanliness</td>

<td></td><td></td><td>m B992</td><td>B992</td><td>m B992</td><td>B992</td>

<td></td><td></td><td>(%)</td><td>(% A / a)</td><td>(%)</td><td>(% A / a)</td>

<td>1198-CB-014</td><td>Tin sulphate (II)</td><td></td><td></td><td></td><td></td>

<td>1198-CB-0881</td><td>Tin oxide (II)</td><td>1.50</td><td>94.53</td><td>0.64</td><td>99.36</td>

<td>1198-CB-088</td><td>Tin bromide (II)</td><td>34.04</td><td>73,09</td><td>9.45</td><td>91.36</td>

<td>2</td><td></td><td></td><td></td><td></td><td></td>

<td>1198-CB-0883</td><td>Tin oxalate (II)</td><td>51.53</td><td>65.72</td><td>57.04</td><td>62.85</td>

<td>1198-CB-0884</td><td>Tin sulphate (II)</td><td>Lack reaction</td><td>0.00</td><td>Lack reaction</td><td>0.00</td>

<td>1198-CB-0885</td><td>Pyrophosphate hydrate tin (II)</td><td>296.65</td><td>24,48</td><td>241.01</td><td>28,43</td>

<td>1198-CB-0886</td><td>Tin (II) ethylhexanoate</td><td>0.29</td><td>99.71</td><td>1.17</td><td>98.84</td>

<td>1198-CB-0887</td><td>Methanesulfonic acid tin (II) (50% solution in water)</td><td>4,601.42</td><td>2.11</td><td>5,540.91</td><td>1.76</td>

<td>1198-CB-0888</td><td>triflates n tin (II)</td><td>2,442.82</td><td>3.90</td><td>2,442.28</td><td>3.89</td>

[0062] Based on the relative completion of these reactions, tin (II) bromide, tin (II) oxalate and tin (II) ethylhexanoate were selected for further testing for the control of tin (II) oxide. The tin input reactions were scaled to 5 g to prepare enough material for total analysis. Reactions were left for 96 hours instead of 48 due to the weekend. The results are shown in Table 7.

<td colspan="6">TABLE 7: IPC RESULTS, ALTERNATIVE SALT IN THE LOW-TEMPERATURE METHOD</td>

<td>Experience</td><td>Salts tested tin</td><td colspan="2">24 hours</td><td colspan="2">96 hours</td>

<td></td><td></td><td>B991 terms of B992</td><td>The peak cleanliness B992 (% a / a)</td><td>B991 terms of B992</td><td>The peak cleanliness B992 (% a / a)</td>

<td>1198-CB-091</td><td>Tin oxide (II)</td><td>8.2</td><td>91.55</td><td>3.5</td><td>96.6</td>

<td></td><td>(CONTROL)</td><td></td><td></td><td></td><td></td>

<td>1198-CB-092</td><td>Tin bromide (II)</td><td>222.7</td><td>30.95</td><td>2841</td><td>3.40</td>

<td>1198-CB-093</td><td>Tin oxalate (II)</td><td>3360</td><td>2.9</td><td>986.9</td><td>9.2</td>

<td>1198-CB-094</td><td>ethylhexanoate tin (II)</td><td>6.6</td><td>93.6</td><td>5.7</td><td>94.6</td>

[0063] Due to incomplete reactions, 1198-CB-092 and 1198-CB-093 were discarded after 96 hours. Reactions 1198-CB-091 and 1198-CB-094 were isolated according to the procedure. In both cases, the yield observed after drying was 79.7% (4.7 g). Both products were subjected to the last purification process (RPA438-04-ES) and the isolated products were analyzed. Tin oxide (II) (1198-CB-091) and tin (II) ethylhexanoate salts gave products with 99.2% and 99.4% purity respectively (Table 8).

<td colspan="5">TABLE 8: ANALYTICAL RESULTS, ALTERNATIVE SALT IN THE LOW-TEMPERATURE METHOD</td>

<td rowspan="2">Test</td><td colspan="2">Raw Products</td><td colspan="2">Purified Products</td>

<td>1198CB-0961</td><td>1198-CB-0971</td><td>1198-CB-0981</td><td>1198-CB-0991</td>

<td>Experience</td><td>Oxide</td><td>ethylhexanoate</td><td>Oxide</td><td>ethylhexanoate</td>

<td>HPLC purity (% a / a)</td><td>98.8</td><td>99.3</td><td>99.2</td><td>99.4</td>

<td>HPLC quantity tested (current% w / w)</td><td>95</td><td>100</td><td>101</td><td>100</td>

<td>Description</td><td>ND</td><td>ND</td><td>compatible</td><td>compatible</td>

<td>IR identification</td><td>ND</td><td>ND</td><td>compatible</td><td>compatible</td>

<td>HPLC identification</td><td>ND</td><td>ND</td><td>compatible</td><td>compatible</td>

<td>KF (% w / w)</td><td>ND</td><td>ND</td><td>&lt;0.05</td><td>&lt;0.05</td>

<td>residues solvents: acetic and formic (% w / w)</td><td>ND</td><td>ND</td><td>&lt;0.1</td><td>&lt;0.1</td>

<td>Solubility</td><td>ND</td><td>ND</td><td>Soluble</td><td>Soluble</td>

EXAMPLE 5 [0064] In process RPA438-01-ES, 5% palladium on carbon (50% wet weight, 0.6 kg) is hydrogenated in formic acid (60 L) under an inert atmosphere of 40 ° C / (60 to 65 psi) ) for a period of 12 hours. After cooling, hemin (B990, 6 kg) is added to the reaction vessel as a suspension in formic acid (60 L). The hemin is then hydrogenated at 85 to 90 ° C / 60 psi for 1 to 2 hours and then hydrogenated at 45 to 50 ° C / 60 psi for a further 24 to 36 hours. The reaction is monitored using HPLC. Upon completion, the reaction is cooled to 20 to 25 ° C, Darco KB-G is introduced and Hyflo Supercel is filtered to remove the catalyst. After filtering the palladium catalyst, the reaction mixture was divided into three portions equal in mass.

[0065] With reference to FIG. 1, one third of the reaction mixture was isolated as CK1 in the RPA438-01-ES process, which was then carried out for the remainder of the process (1165-CB-155-1). The filtration solution is concentrated by vacuum distillation to a residual volume and 30 L of methyl tert-butyl ether (MTBE, 120 L) is added to the concentrate for at least 1 hour to precipitate the intermediate. The resulting suspension is cooled to -20 to -25 ° C over 1 hour and stirred for 4 hours after filtration. The mass is vacuum dried to remove MTBE residue, yielding 5 to 6 kg of mesoporphyrin IX formate (CK1, 85 to 100% yield).

One third was isolated directly as B991 in the RPA438-02-ES process and was followed for the remainder of the process (1165-CB-154-1). In RPA438-02ES, mesoporphyrin IX formate (CK1, 5 kg as the free base) is dissolved in formic acid (22 L) and treated with activated charcoal (Darco KBG, 0.2 kg) and Hyflo Supercel (0.4 kg) and stirred for 2 hours at 20 to 25 ° C before filtering to remove solids. The filtrate is concentrated using vacuum distillation to a residual volume of 12 L. To this solution is added 1N HCl solution (13 L) for at least 1 hour to precipitate the intermediate. The resulting suspension is stirred at 20 to 25 ° C under nitrogen for 2 hours before filtration. After filtration the product is dried under a stream of nitrogen to obtain 3.4 to 4,

[0067] The remaining third part of the reaction mixture can be transferred directly to the tin introduction reaction, RPA438-03-ES, as a filtered solution of formic acid to give Stannsoporfin (B992) (1165-CB-153-1). In process RPA43803-ES, tin (II) oxide powder (SnO, 1.7 kg) was suspended in 40.5 L of acetic acid at 20 to 25 ° C and then heated to from 60 to 65 ° C under nitrogen. To this suspension is added a solution of mesoporphyrin IX dihydrochloride (B991, 2.1 kg) in 10.5L formic acid for 6 hours. The reaction was carried out under nitrogen at 60 to 65 ° C for at least 12 hours. The reaction is monitored using HPLC. After completion of the reaction, 17 L of water was added over 0.5 hour. The reaction mixture is then cooled to from about 20 ° C to about 25 ° C for about 0.5 hour, stirred for about 1 to about 3 hours and then filtered. The filtration mass is washed with distilled water (USP), suspended in 1N HCl and heated to 85 to 95 ° C for 1 to 3 hours. The suspension is then cooled to 20 to 25 ° C, stirred for 0.5 hour, filtered, washed with distilled water (USP) and dried in nitrogen to give 1.3 to 1.5 kg of raw tin mesoporphyrin IX dichloride (StannsoporfinTM, B992) (efficiency from 65 to 75%).

[0068] All B992 products obtained were also subjected to final purification, RPA438-04-ES. In the RPA438-04-ES process, the crude tin mesoporphyrin IX dichloride (B992, 1.7 kg) was dissolved in 2% ammonium hydroxide (22 L). A pH measurement was carried out to ensure that the pH was 2: 9.0. The solution was processed using Darco KB-G (0.1 kg) and Hyflo Supercel (0.2 kg), stirred for 1 to 2 hours and filtered to remove solids. The filtrate is then added dropwise to acetic acid (44 L) containing hydrochloric acid (31%, 2.7 L) while maintaining the temperature at 20 to 25 ° C. PH measurement is again performed to ensure pH &lt;1.0. The resulting suspension is stirred for 1 to 2 hours under nitrogen to isolate the product by filtration. The wet mass is then triturated in 3N HCl (35 L) at 85 to 90 ° C and stirred for 16 to 18 hours to convert the crystalline form to a monomer. The suspension is cooled to 20 to 25 ° C and the product is isolated using filtration. The product is washed using 0.3 N HCl (16 L) and dried under a stream of nitrogen to obtain 1.2 to 1.6 kg of stannsoporfin API (yield 70 to 90%). The results are summarized in Table 9.

TABLE 9: ANALYTICAL RESULTS, COMBINATION OF PROCESS STAGES

<td>Test</td><td colspan="4">1165-CB-153-1</td><td colspan="4">1165-CB-1541</td><td colspan="4">1165-CB-1551</td>

<td>experienced e</td><td colspan="4">No isolates</td><td colspan="4">Isolate B991</td><td colspan="4">isolate B991 / CK1 (Control)</td>

<td>Efficiency:</td><td colspan="4">61.9</td><td colspan="4">54.2</td><td colspan="4">48.2</td>

<td rowspan="8">Cleanliness with HPLC (% a / a)</td><td colspan="4">99,3</td><td colspan="4">99.5</td><td colspan="4">99.6</td>

<td rowspan="7"></td><td>RRT</td><td>% A / a</td><td rowspan="7"></td><td rowspan="6"></td><td>RR t</td><td>%and/ and</td><td rowspan="6"></td><td rowspan="5"></td><td>RR t</td><td>%and/ and</td><td rowspan="5"></td>

<td>0.22</td><td>0.06</td><td>0.0 7</td><td>0.06</td><td>0.8 8</td><td>0.05</td>

<td>0.67</td><td>0.07</td><td>0.8 8</td><td>0.05</td><td>0.9 1</td><td>0.03</td>

<td>0.88</td><td>0.09</td><td>0.9 1</td><td>0.03</td><td>0.9 5</td><td>0.03</td>

<td>0.91</td><td>0.06</td><td>0.9 5</td><td>0.03</td><td>Inn e</td><td>&lt;0 05</td>

<td>0.95</td><td>0.02</td><td>Inn e</td><td>&lt;0 05</td><td colspan="4" rowspan="2"></td>

<td>Other</td><td>&lt;0.05</td><td colspan="4"></td>

<td>HPLC test (% W / w)</td><td colspan="4">96.8</td><td colspan="4">98.2</td><td colspan="4">99.5</td>

<td>Description</td><td colspan="4">Compatible</td><td colspan="4">Compatible</td><td colspan="4">Compatible</td>

<td>Identification IR</td><td colspan="4">compatible</td><td colspan="4">compatible</td><td colspan="4">compatible</td>

<td>Identification HPLC</td><td colspan="4">compatible</td><td colspan="4">compatible</td><td colspan="4">compatible</td>

<td>KF (% w / w)</td><td colspan="4">1.2</td><td colspan="4">0.4</td><td colspan="4">0.6</td>

<td>Residues of solvents (acetone)</td><td colspan="4">&lt;500</td><td colspan="4">&lt;500</td><td colspan="4">&lt;500</td>

<td>Residues of Solvents (Octane and Ants)</td><td colspan="4">0.1</td><td colspan="4"></td><td colspan="4"></td>

<td>Iron (ppm)</td><td>6.55</td><td>2.94</td><td>2.84</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Palladium (ppm)</td><td>6.95</td><td>&lt;2</td><td>&lt;2</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>soluble bone</td><td>Dissolution Stop Limit Current</td><td>Dissolution Stop Limit Current</td><td>Dissolution Stop Limit Current</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Chloride (% w / w)</td><td>7.3</td><td>9.69</td><td>9.97</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

[0069] As can be seen from the table, each additional insulation reduces the overall efficiency of the process, increasing the test amount and purity of the final product. All three materials underwent a qualification other than HPLC. The final purification method can increase the amount of raw API tested. Each of these materials was subjected to final purification, RPA438-04-ES, up to the second iteration and was re-analyzed using HPLC for purity and the amount tested (Table 10).

<td colspan="4">TABLE 10: ANALYTICAL RESULTS, COMBINATION OF PROCESS STAGES ADDITIONAL CLEANSING</td>

<td>Test</td><td>1198-CB-020-1</td><td>1198-CB-021-1</td><td>1198-CB-022-1 (CONTROL)</td>

<td>Output Material</td><td>1165-CB-153-1</td><td>1165-CB-154-1</td><td>1165-CB-155-1</td>

<td>Efficiency:</td><td>92.0</td><td>92.0</td><td>94.0</td>

<td rowspan="2"></td><td rowspan="2"></td><td rowspan="2"></td><td></td>

<td>RRt% a / a</td>

<td rowspan="9"></td><td rowspan="11"></td><td rowspan="2">RRT</td><td rowspan="2">% A / a</td><td rowspan="11"></td><td rowspan="10"></td><td rowspan="2">RRT</td><td rowspan="2">% A / a</td><td rowspan="10"></td><td rowspan="11"></td><td></td><td></td><td rowspan="11"></td>

<td rowspan="2">0.22</td><td rowspan="2">0.05</td>

<td rowspan="2">0.22</td><td rowspan="2">0.07</td><td rowspan="2">0.07</td><td rowspan="2">0.06</td>

<td rowspan="2">0.88</td><td rowspan="2">0.05</td>

<td rowspan="2">0.39</td><td rowspan="2">0.05</td><td rowspan="2">0.14</td><td rowspan="2">0.05</td>

<td rowspan="2">0.95</td><td rowspan="2">0.05</td>

<td rowspan="2">0.44</td><td rowspan="2">0.05</td><td rowspan="2">0.22</td><td rowspan="2">0.05</td>

<td rowspan="2">1.08</td><td rowspan="2">0.13</td>

<td rowspan="2">0.64</td><td rowspan="2">0.05</td><td rowspan="2">Other</td><td rowspan="2">&lt;0.05</td>

<td rowspan="2">HPLC purity (% a / a)</td><td rowspan="2">Other</td><td rowspan="2">&lt;0.05</td>

<td>Other</td><td>&lt;0.05</td><td colspan="4"></td>

<td>HPLC quantity tested (% w / w)</td><td colspan="4">98 (as it is)</td><td colspan="4">98 (as it is)</td><td colspan="4">99 (as it is)</td>

[0001] With the exception of tin sulfate, all tin salts were selected for stannsoporfin use in the tin introduction reaction using both the oxidation and non-oxidation methods. It is believed that higher purity can be achieved with all such tin salts. Using ethylhexanoate as a stannous carrier, it was possible to obtain stannsoporfin equal in quality to the product that was obtained using tin oxide.