Use of peptide compounds for treating bone cancer pain, chemotherapy- and nucleoside-induced pain

The present invention relates to the use of a class of peptide compounds for the treatment of bone cancer-related pain, for the treatment of chemotherapy-induced pain, and for the treatment of nucleoside-induced pain as defined in the claims.

[0002] It is known that certain peptides have activity against the central nervous system (CNS) and are useful for the treatment of epilepsy and other CNS disorders. These peptides, which are described in US Patent No. 5,378,729, have formula (Ia):

R-NH

<img file="PL1781276T3_D0001.tif" />

II

II

Ra formula (Ia) in which

R is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, aryl, aryl lower alkyl, heterocyclic, heterocyclyl lower alkyl, lower alkyl heterocyclic, lower cycloalkyl, lower cycloalkyl lower alkyl, and R is unsubstituted or is substituted by at least one electron withdrawing group or electron donating group;

R₁ represents a hydrogen atom or a lower alkyl, lower alkenyl, lower alkynyl, aryl lower alkyl, aryl, heterocyclyl lower alkyl, heterocyclic, lower cycloalkyl, lower cycloalkyl lower alkyl group, each of which is unsubstituted or substituted by an electron donating group or electron withdrawing group; and

R₂ and R₃ are independently hydrogen, lower alkyl, lower alkenyl, lower alkynyl, aryl lower alkyl, aryl, heterocyclic, heterocyclyl lower alkyl, lower alkyl heterocyclic, lower cycloalkyl, lower cycloalkyl lower alkyl, or ZY, where R₂ and R₃ may be unsubstituted or substituted by at least one electron withdrawing group or electron donating group;

Z means O, S, S (O)_and, NO₄, PR₄ or chemical bond;

Y is hydrogen, lower alkyl, aryl, lower alkyl, lower alkenyl, lower alkynyl, halogen, heterocyclic, heterocyclyl lower alkyl, and Y may be unsubstituted or substituted by an electron donating group or electron withdrawing group, provided that when Y is halogen, Z is a chemical bond, or

Taken together, ZY means NR₄NO₅R₇, NO₄OR₅, ONR₄R₇, OPR₄R₅, PR₄OR₅, SNR₄R₇, NR4SR7, SPR4R5 or PR4SR7, NR4PR5R6 or PR4NR5R7,

NR4C-R5, SCRs, NR4C-OR5,

II

II

SC-ORb;

II

R₄, R₅ and R₆ are independently hydrogen, lower alkyl, aryl, lower aryl, lower alkenyl, or lower alkynyl, where R₄, R₅and R₆ may be unsubstituted or substituted by an electron withdrawing group or an electron donating group; and

EP 1 781 276 B1

R₇ means R₆ or COOP₈ or COR₈;

R₈ is a hydrogen atom or a lower alkyl or aryl lower alkyl group, and the aryl or alkyl group may be unsubstituted or substituted by an electron withdrawing group or an electron donating group; in is 1-4; and a is 1-3.

[0003] US Patent No. 5,773,475 also discloses additional compounds useful for the treatment of CNS disorders. These compounds are N-benzyl-2-amino-3-methoxy-propionamides having formula (IIa):

Η Η Η ι 1 I

Ar-CHr-NCCNCR, 'Μ II ο ch₂ ο

AND

Rs formula (IIa) in which

Ar is an aryl group which is unsubstituted or substituted by a halogen atom; R₃is a lower alkoxy group; and R₁ is a methyl group.

[0004] None of these patents, however, describes the use of these compounds for the treatment of cancer-related pain, in particular bone cancer-related pain, for the treatment of chemotherapy-induced pain and for the treatment of nucleoside-induced pain as defined in the present claims.

[0005] WO 02/074297 relates to the use of a compound of formula (IIa), wherein Ar is a phenyl group which may be substituted by at least one halogen atom, R₃ is a lower alkoxy group containing 1-3 carbon atoms and R₁ is a methyl group, for the preparation of pharmaceutical compositions useful for the treatment of allodynia associated with peripheral neuropathic pain.

[0006] WO 02/074784 relates to the use of a compound of formula (Ia) or / and formula (IIa) which exhibits anti-nociceptive (anti-harmful) stimuli for the treatment of various types and symptoms of acute and chronic pain, especially non-neuropathic pain associated with inflammation, e.g., pain associated with rheumatoid arthritis or / and secondary pain associated with osteoarthritis.

[0007] Currently, there is no analgesic that would have a highly potent effect on various pain syndromes. Various mechanisms leading to inflammatory or neuropathic pain make it difficult to identify compounds that have general analgesic activity. We are only at the beginning of understanding the mechanisms behind various pain syndromes, such as cancer-related pain (e.g. tumor-induced pain associated with bone cancer), chemotherapy-induced pain or nucleoside-induced pain, which all appear to have a variety of molecular origins. Antidepressants, anticonvulsants or synthetic opiates, which describe groups of compounds used to treat pain, do not have a common pattern of efficacy in the treatment of pain syndromes. It makes it harder

EP 1 781 276 B1 predicts the activity of new compounds in various pain syndromes and requires detailed characterization in numerous animal models.

[0008] Neuropathic pain after trauma or dysfunction of the peripheral or central nervous system remains a difficult clinical problem for which there are no effective treatments (Bennett, 1994; Murphy and Reid, 2001). Anticonvulsants are used to control certain forms of neuropathic pain (Sindrup and Jenssen, 1999; Jensen, 2002). SPM 927 (R-2-acetamido-N-benzyl-3-methoxypropionamide) also called harkoseride or ADD 234037 is a new anticonvulsant. It belongs to a series of functionalized amino acids that have been synthesized as a new class of anticonvulsants (Kohn et al. 1991).

WO 02/15922 discloses the use of SPM 927 for the treatment of neuropathic pain. Current studies show the analgesic effect of SPM 927 in rat models of cancer-related pain, in particular bone cancer-related pain, chemotherapy-induced pain, and nucleoside analogue-induced pain.

[0009] Bone is the third most common metastatic site after lungs and liver, and the main site for metastatic disease in patients with breast, prostate and lung cancer. Bone disease lesions that are the result of metastatic disease also cause severe bone pain, which is a major clinical problem in cancer patients. This type of pain is difficult to treat because of its intermittent, progressive nature, and its exacerbation by movement. The main symptom in this model of pain is mechanical allodynia. Thermal hyperalgesia and mechanical hyperalgesia have also been shown as measured by the weight-bearing difference in two hind limbs (Medhurst et al., 2002). Treatment of bone pain in human patients is largely limited to the use of synthetic opiates, however, the effectiveness of strong synthetic opiates is minimal, and effective doses give a number of debilitating side effects. Consequently, there is a clinical need for new therapies that can be used to prevent, treat and alleviate tumor-induced bone pain. Candidate therapies for cancer-induced bone pain treatment can be evaluated using a rat model, as the rat is better suited to test behavioral responses to pain stimuli. One model involves injecting rat mammary gland cancer cells into the proximal bone marrow space using the pain assessment endpoint (Medhurst et al., 2002), which was performed on days 7 to 15 after tumor implantation.

[0010] Chemotherapy-induced pain is a form of neuropathic pain associated with neurotoxic drugs such as vinca alkaloids, e.g. vincristine, and is characterized by painful paresthesia and abnormal sensations. The clinical efficacy of the anti-cancer drug vincristine is limited by the development of mixed sensorimotor neuropathy (Casey et al., 1973, Tanner et al., 1998 et al. 1998), which appears to occur in two main stages (Weiss et al., 1974 ). In the early stages, peripheral axons are destroyed by vincristine, and the main symptoms are paresthesia and abnormal sensations. In the late stage, which occurs more often when higher doses are administered for a longer period of time, axons die and the main clinical finding is loss of motor function. The rat vincristine model described seems to reflect the early stage of vincristine-induced chemotherapeutic neuropathy. Although the underlying mechanism is not yet fully understood, it has been reported to cause the disorganization of the cytotope microtubular axons as well as the increase in cross-section of non-myelin sensory axons (Quasthoff et al., 2002). These results showed that changes in the structure of microtubules in sensory neurons receiving harmful stimuli accompany vincristine induced hyperalgesia.

[0011] Painful peripheral neuropathy induced by nucleoside analogues is becoming increasingly recognized as an important source of morbidity in people infected with human immunodeficiency virus (HIV) (Cohen, 2002). This severely debilitating side effect may force you to shorten or even discontinue AIDS (acquired immunodeficiency syndrome) therapy (Yatvin et al., 1999). This neuropathy is characterized by a sudden onset of about 10 weeks of intensive burning treatment in both hands-saving feet, which reaches severe intensity over a period of days (Dubinsky et al., 1989). The biochemical mechanism underlying this neuropathy remains to be explained, although mitochondrial toxicity has been reported to contribute to the development of this neuropathy. Recently, it has been reported that poisoning rats with anti-retroviral nucleoside analogues for the treatment of AIDS (ddC (2 ', 3'-dideoxycytidine), ddl (2', 3'-didezoxyinosine) or d4T (2 ', 3'-didehydro-3'- (deoxythymidine)), it gives increased reception of harmful stimuli in the rat (Joseph et al., 2004). The mechanism involved appears to be different from that of other metabolic or toxic painful peripheral neuropathy models, since anti-hyperalgesia drugs effective in these models, protein kinase A inhibitors, protein kinase C, protein kinase G, mitogen activated protein kinase p42 / p44 (ERK1 / 2) and nitrogen oxygen synthase do not affect peripheral neuropathies, and do not affect the hypersensitivity induced by reverse transcriptase nucleoside inhibitors. The only means capable of reversing this hypersensitivity in poisoned animals are intercellular calcium modulators (TMB-8 and Quin-2), which strongly suggests the role of intercellular calcium in this type of neuropathic pain.

[0012] Chemotherapy is used to treat patients suffering from cancer and HIV, e.g. treatment with vinca alkaloids such as vincristine or taxol, suramin, cisplatin, carboplatin or oxaliplatin. In addition, patients suffering from HIV and / or cancer are also treated with antiretroviral or antiviral drugs.

[0013] The use of compounds of formula (IIb) for the treatment of bone cancer-related pain, for the treatment of chemotherapy-induced pain and for the treatment of nucleoside-induced pain as defined in the claims has not yet been described. Thus, the present invention relates to the use of said compounds of formula (IIb) for the preparation of a pharmaceutical composition for the prevention, alleviation and / or treatment of pain associated with bone cancer. The present invention furthermore relates to the use of compounds of formula (IIb) for the preparation of a pharmaceutical composition for the prevention, alleviation and / or treatment of chemotherapy-induced pain, such as chemotherapy-induced neuropathic pain, vinca alkaloid-induced pain, vincristine-induced pain and / or pain induced by taxol, suramin, cisplatin, carboplatin or / and oxaliplatin. The present invention furthermore relates to the use of compounds of formula (IIb) for the preparation of a pharmaceutical composition for the prevention, alleviation and / or treatment of pain induced by nucleosides and / or nucleoside analogues selected from pain induced by anticancer or / and antiviral nucleoside analogues and / or induced pain by antiviral analogies

Nucleosides in the treatment of AIDS and / or pain induced by AZT (3'-azidothymidine), ddC, ddl or / and d4T.

[0014] The invention also relates to the use of compounds of formula (IIb) for the preparation of a pharmaceutical composition for preventing, alleviating and / or treating bone cancer pain, chemotherapy induced pain, and / or pain induced by at least one nucleoside or / and at least one nucleoside analog as defined in the claims.

[0015] Surprisingly, the use of compounds of compounds (IIb), especially (R) -2-acetamido-N-benzyl-3-methoxypropionamide (SPM 927) reduced mechanical and thermal hyperalgesia as well as mechanical and thermal allodynia in a model of tumor-induced pain associated with bone cancer, in the model of neuropathic pain induced by chemotherapy and in the model of pain induced by nucleoside analogues.

[0016] The compound for use according to the present invention has the general formula (IIb)₂AND

R-NH - [- C-CN H-1 n'-C-Ri

II I II o I o

Ra formula (Ilb) in which

Ar is a phenyl group which is unsubstituted or substituted by at least one halogen atom,

R₃ means CH₂-Q, where Q is a lower alkoxy group containing 1-3 carbon atoms, and R₁ is a lower alkyl group having 1-3 carbon atoms.

[0017] The present invention also provides a pharmaceutical composition comprising a compound of formula (IIb) for use in the prevention, alleviation or / and treatment of pain associated with bone cancer. The present invention further provides a pharmaceutical composition comprising a compound of formula (IIb) for use in preventing, alleviating or / and treating chemotherapy-induced pain, such as chemotherapy-induced neuropathic pain, vinca alkaloid-induced pain, vincristine-induced pain and / or pain induced by taxol, suramin, cisplatin, carboplatin or / and oxaliplatin. The present invention furthermore relates to a pharmaceutical composition comprising a compound of formula (IIb) for use in the prevention, alleviation or / and treatment of pain induced by nucleosides or / and nucleoside analogues selected from anticancer or / and antiviral nucleoside analogues and / or pain induced by nucleoside analogues antiviral in the treatment of AIDS, and / or pain induced by AZT, ddC, ddl or / and d4T.

[0018] The invention also provides a pharmaceutical composition comprising a compound of formula (IIb) for use in preventing, alleviating and / or treating bone cancer pain, chemotherapy-induced pain, and / or pain induced by at least one nucleoside or / and at least one nucleoside analog as defined in the claims.

[0019] "Lower alkyl" groups, used alone or in combination with other groups, mean a lower alkyl group having 1 to 3 carbon atoms, and may be straight or branched chain. These groups include methyl, ethyl, propyl and isopropyl.

[0020] "Lower alkoxy" groups mean a lower alkoxy group containing from 1 to 3 carbon atoms, and may be straight or branched chain. These groups include methoxy, ethoxy and propoxy.

[0021] The term "halogen" includes fluorine, chlorine, bromine and iodine.

[0022] R₁ is a lower alkyl group. Most preferably the R group₁ is a methyl group.

[0023] Most preferably R₃ means CH₂-Q where Q is a methoxy group.

[0024] The most preferred compounds include: (R) -2-acetamido-N-benzyl-3-methoxy-propionamide; O-methyl-N-acetyl-D-serine-m-fluorobenzyl-amide; O-methyl-N-acetyl-D-serine-p-fluorobenzyl-amide;

[0025] It should be understood that various combinations and permutations of the Markusch R groups described herein are contemplated to be within the scope of this invention.₁, R₂, R₃, R. In addition, the present invention also encompasses compounds and compositions that contain one or more elements of each of Markusha moieties in R₁, R₂, R₃, and R, and their various combinations. Thus, for example, the present invention contemplates that R₁ it may be one or more substituents given above in combination with any and all of R₂, R₃, and R.

[0026] The compounds used in the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. The configuration around each of the asymmetric carbon atoms can be in the form of either D or L. It is known in the art that the configuration around chiral carbon atoms can also be described as R or S in the Cahn-Prelog-Ingold nomenclature system. The present invention contemplates all of the various configurations around each of the asymmetric carbon atoms, including various enantiomers and diastereomers, as well as racemic mixtures and mixtures of enantiomers, diastereomers, or both.

[0027] In the main chain there is asymmetry on the carbon atom to which the R groups are attached₂ and R₃. The compounds of the present invention have the formula

<img file="PL1781276T3_D0002.tif" />

in which R, R₁, R₂, R₃ have the meanings defined herein. R₂ is hydrogen. R is a benzyl group in which its phenyl ring is unsubstituted or substituted by at least one halogen atom.

[0028] The term configuration as used herein is intended to refer to the configuration around the carbon atom to which R is attached₂ and R₃, even though other chiral centers may be present in the molecule. Thus, it should be understood that reference to a particular configuration, such as D or L, is intended to mean the D or L stereoisomer on the carbon atom to which R is attached₂ and R₃. However, this also includes all possible enantiomers and diastereomers on other chiral centers, if such are present.

[0029] The compounds for use in the present invention include all optical isomers, i.e., the compounds of the present invention are either the L stereoisomer or the D stereoisomer (on the carbon atom to which R is attached₂ and R₃). These stereoisomers may be present in mixtures of L and D stereoisomers, e.g., in racemic mixtures. The stereoisomer D is preferred.

[0030] More preferred is the compound of formula (III) in the R configuration, preferably substantially enantiomerically pure, wherein R is a benzyl group which is unsubstituted or substituted by at least one halogen atom in which R₃ means CH₂-Q where Q is a lower alkoxy group containing 1-3 carbon atoms, and in which R₁is a methyl group in which R₂ is H. Preferably R is an unsubstituted benzyl group or a benzyl group substituted by at least one halogen atom which is a fluorine atom.

[0031] Depending on the substituents, the present compounds may also form addition salts. It is contemplated that all of these forms, including mixtures of stereoisomeric forms, are within the scope of this invention.

[0032] The preparation of the compounds used is described in US Patent Nos. 5,378,729 and 5,773,475.

[0033] The compounds used in the present invention are useful as such as represented by formula (IIb) or may be used in the form of salts due to their basic nature due to the presence of a free amino group. Thus, the compounds of formula (IIb) form salts with various inorganic and organic acids, including pharmaceutically acceptable acids. Salts with therapeutically acceptable acids are obviously useful in the preparation of the formulation, where the increased water solubility is most preferred.

[0034] These pharmaceutically acceptable salts also have therapeutic efficacy. These salts include the salts of inorganic acids such as hydrochloric, hydroiodic, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, as well as salts of organic acids such as tartaric, acetic, citric, malic, benzoic, perchloric, glycolic, gluconic, succinic, arylsulfonic (e.g., p-toluenesulfonic, benzenesulfonic acid), phosphoric and malonic acid.

[0035] Preferably the compound used in the present invention is used in therapeutically effective amounts.

[0036] The physician will determine the dosage of the present therapeutic agents that will be most useful and will vary depending on the form of administration and the particular compound selected, and will also vary depending on the patient being treated, the age of the patient, the type of disease being treated. Generally, he will want to start treatment at low dosages substantially less than the optimal dose of the compound and increase the dosage by small increments until optimal effect is achieved under the circumstances. When the composition is administered orally, larger amounts of active agent will be needed to achieve the same effect as for a smaller amount administered parenterally. The compounds are useful in the same way as comparable therapeutic agents and the dosage level is of the same order of magnitude that is generally used with these other therapeutic agents.

[0037] In a preferred embodiment, the compounds of the present invention are administered in amounts ranging from 1 mg to 100 mg per kilogram body weight per day, more preferably in amounts ranging from 1 mg to 10 mg per kilogram body weight for day. This dosage regimen can be adjusted by the physician for an optimal therapeutic response. Patients in need thereof may be treated with doses of the compound of the present invention of at least 50 mg / day, preferably of at least 200 mg / day, more preferably of at least 300 mg / day, and most preferably of at least 400 mg / day. Generally, a patient in need thereof may be treated with doses of at most 6 g / day, more preferably at most 1 g / day, and most preferably at most 600 mg / day. However, in some cases higher or lower doses may be needed.

[0038] In another preferred embodiment, the daily doses are increased until the scheduled daily dose is reached, which is maintained during further treatment.

[0039] In yet another preferred embodiment, several divided doses may be administered per day. For example, three doses per day, preferably two doses per day, may be administered. It is more preferable to administer a single dose per day.

[0040] In yet another preferred embodiment, an amount of compounds of the present invention can be administered that results in a plasma concentration of 0.1 to 15 μg / ml (at the lowest point) and 5 to 18.5 μg / ml (at the highest point point), calculated as the average for many treated.

[0041] The compounds of formula (IIb) can be administered in a convenient manner such as oral, intravenous (when they are water-soluble), intramuscular, intrathecal or subcutaneous routes. Oral and / or intravenous administration is preferred.

[0042] The pharmaceutical composition for use in the present invention may be prepared for a treatment regimen as described above, in particular for treatment with doses as described above, for obtaining plasma concentrations as described above, for administration periods and / or routes of administration as specified in embodiments of the present invention as described above.

[0043] The compound for use in the present invention may be administered in combination with the administration of a further active agent for the prevention, amelioration and / or treatment of a viral infection such as retroviral infection, HIV infection including AIDS, cancer such as breast cancer, prostate cancer, cancer lungs, bone cancer, metastatic disease, and / or tumor progression through infiltration or pressure on the bone, viscera, soft tissue or nerves. The compound for use in the present invention and the further active agent may be administered together, i.e. as a single dose, or may be administered separately, i.e. as a separate dose. Thus, the pharmaceutical composition for use in the present invention may contain a compound of the present invention as defined above and may further comprise a further active agent for preventing, ameliorating or / and treating a viral infection such as a retroviral infection, HIV infection including AIDS, such a cancer like breast cancer, prostate cancer, lung cancer, bone cancer, metastatic disease, and / or tumor progression through infiltration or pressure on the bone, viscera, soft tissue or nerves. The pharmaceutical composition may be in a single dose form or may be in separate doses comprising a first composition comprising a compound of the present invention as defined above and a second composition comprising a further active agent.

[0044] The compounds for use in the present invention may be used in the preparation of a pharmaceutical composition as described above.

[0045] The compounds of formula (IIb) may be administered orally, for example, with an inert diluent or with an absorbable edible carrier, or they may be enclosed in hard or flexible gelatin capsules, or they may be compressed into tablets, or may be incorporated directly to the food diet. For oral therapeutic administration, the active compound of formula (IIb) may be combined with excipients and used in the form of swallow tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups and wafers. Such compositions and preparations should contain at least 1% of active compound of formula (IIb). The percentage in the compositions and preparations can of course be varied and may conveniently be between 5 to 80% of the weight of the unit. The amount of active compound of formula (IIb) in such therapeutically useful compositions is such that a useful dosage will be obtained. Preferred compositions or preparations according to the present invention contain between 10 mg and 6 g of the active compound of formula (IIb).

[0046] Tablets, troches, pills, and capsules may also contain the following: a binder such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch and alginic acid, a lubricant such as magnesium stearate; and a sweetener such as sucrose, lactose or saccharin or a flavoring such as peppermint oil, wintergreen oil, or cherry flavor may be added. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier.

[0047] Various other materials may be present as coatings or otherwise modify the physical form of the dosage unit. For example, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and a flavoring such as cherry or orange flavor. Of course, any material used to make any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts used. In addition, the active compound may be incorporated into sustained-release preparations and formulations. For example, sustained release dosage forms are contemplated where the active ingredient is bound to an ion exchange resin, which may optionally be coated with a diffusion barrier coating to modify the resin's release properties.

[0048] The active compound may also be administered parenterally or intraperitoneally. Dispersions can also be made in glycerin, liquid polyethylene glycols, and mixtures thereof, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0049] Pharmaceutical forms suitable for injection include sterile aqueous solutions (when the active compound is water-soluble) or dispersions and sterile powders for freehand preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms such as bacteria

EP 1 781 276 B1 and mushrooms. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), useful mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be achieved using a variety of antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and thimerosal. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be achieved by using absorption delaying agents in the compositions, for example, aluminum monostearate and gelatin.

[0050] Sterile injectable solutions are prepared by containing the active compound in the required amount in a suitable solvent with the various other ingredients listed above as needed, followed by sterilization by filtration. Generally, dispersions are prepared by containing the variously sterilized active ingredient in a sterile vehicle that contains the basic dispersing medium and the other ingredients required from those enumerated above. For sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation are lyophilization techniques with the addition of any additional desired components from their previously sterilized solution by filtration.

[0051] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delaying pharmaceutical active substances as are known in the art. Except when any conventional medium or agent is incompatible with the active ingredient, their use is contemplated in therapeutic compositions. Additional active ingredients may also be included in the composition.

[0052] It is particularly preferred to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units adapted as unitary dosages for the treated mammals; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specificity of the new dosage unit forms is dictated and directly dependent on (a) the specific characteristics of the active material and the specific therapeutic effect to be obtained, and (b) the restrictions specific to the field of creating compositions of such active material for treating the disease in living patients, in a disease condition in which bodily health is impaired as disclosed in detail herein.

[0053] The main active ingredient is combined for convenient and effective administration in effective amounts with a suitable pharmaceutically acceptable carrier in the form of a dosage unit as described previously. The dosage unit form may, for example, contain the main active compound in amounts in the range of 10 mg to 6 g. In proportion, the compound

Active is generally present in an amount of from 1 to 750 mg / ml of carrier. In the case of compositions containing complementary active ingredients, dosages are determined by reference to the usual doses and method of administration of said ingredients.

[0054] The term "patient" or "treated," as used herein, refers to a warm-blooded animal, preferably mammals, such as, for example, cats, dogs, horses, cows, pigs, mice, rats and primates, including humans. The preferred patient is human.

[0055] The term "treat" refers to removing pain associated with a disease or condition or to curing or alleviating a patient's disease or condition.

[0056] The compounds for use in the present invention are administered to a patient suffering from the previously mentioned type disorder in an effective amount. These amounts are equivalent to the therapeutically effective amounts described above.

[0057] The following example demonstrates the properties of SPM 927 in reducing mechanical and thermal hyperalgesia as well as mechanical and thermal allodynia in a model of cancer-induced pain associated with bone cancer, in a model of neuropathic pain induced by chemotherapy and induced by nucleoside analogues.

[0058] The substance used was SPM 927, which is synonymous with the name Harkoseride. According to the normal chemical nomenclature, it is (R) -2-acetamido-N-benzyl-3-methoxypropionamide.

Description of the figures [0059]

Figure 1 describes testing of mechanical allodynia in a bone cancer pain model (in rats). Rats having bone cancer were treated with increasing concentrations of SPM 927 (10 mg, 20 mg and 40 mg) and compared with morphine treated rats having bone cancer, rats having bone cancer without treatment (cells only) and control rats.

Figure 2 describes testing paw heat stimulation on days 14 and 15 in a bone cancer pain model (in rats). Rats having bone cancer were treated with increasing concentrations of SPM 927 (3 mg, 10 mg and 30 mg) and compared with morphine treated rats having bone cancer, rats having bone cancer without treatment (cells only) and control rats.

Figure 3 describes testing of weight loss differences in a bone cancer pain model (rats). Rats having bone cancer were treated with increasing concentrations of SPM 927 (10 mg, 20 mg and 40 mg) and compared with morphine treated rats having bone cancer, rats having bone cancer without treatment (cells only) and control rats.

Figure 4 describes the effect of increasing concentrations of SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) on ​​thermal allodynia in a cold bath test in a chemotherapy-induced pain model (rats treated with vincristine) compared to morphine (3 mg / kg).

Figure 5 describes the effect of increasing concentrations of SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) on ​​heat allodynia in a 38 ° C hot plate test in a model of chemotherapy-induced pain (rats treated with vincristine), compared with morphine (3 mg / kg).

Figure 6 describes the effect of increasing concentrations of SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) on ​​thermal hyperalgesia in a 52 ° C hot plate test in a chemotherapy-induced pain model (vincristine-treated rats), compared with morphine (3 mg / kg).

EP 1 781 276 B1

Figure 7 describes the effect of increasing concentrations of SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) on ​​mechanical hyperalgesia in the paw pressure test in a chemotherapy-induced pain model (rats treated with vincristine) compared to morphine (3 mg / kg).

Figure 8 describes the effect of increasing concentrations of SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) on ​​mechanical allodynia in von Frey filament stimulation tests in a model of chemotherapy-induced pain (rats treated with vincristine) compared to morphine (3 mg / kg).

Figure 9 describes the effect of increasing concentrations of intraperitoneally administered SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) in a heat allodynia test (cold bath) in a model of nucleoside-induced pain (ddC-treated rats), compared to the effect morphine (3 mg / kg administered subcutaneously).

Figure 10 describes the effect of increasing concentrations of intraperitoneally administered SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) in a mechanical allodynia test (brushing at D20) in a nucleoside-induced pain model (ddC-treated rats) compared to by morphine (3 mg / kg administered subcutaneously).

Figure 11 describes the effect of increasing concentrations of intraperitoneally administered SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) in a mechanical allodynia test (von Frey filaments) in a model of pain induced by nucleosides (ddC-treated rats) in compared with morphine (3 mg / kg given subcutaneously).

Figure 12 describes the effect of increasing concentrations of intraperitoneally administered SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) in a hyperalgesia test (hot plate at 52 ° C) in a model of pain induced by nucleosides (ddC-treated rats) , compared with morphine (3 mg / kg administered subcutaneously).

Figure 13 describes the effect of increasing concentrations of intraperitoneally administered SPM 927 (3 mg / kg, 10 mg / kg and 30 mg / kg) in a mechanical hyperalgesia test (paw pressure) in a nucleoside-induced pain model (ddC-treated rats), compared to the effect morphine (3 mg / kg administered subcutaneously).

Example [0060] The effect of systemic administration of SPM 927 was studied in rats in a model of tumor-induced pain associated with bone cancer, in a model of neuropathic pain induced by chemotherapy and induced by nucleoside analogues. SPM 927 reduced mechanical and thermal hyperalgesia as well as mechanical and thermal allodynia in these models. SPM 927 has been shown to be useful as a painkiller for the treatment of pain associated with bone cancer, chemotherapy-induced neuropathy and nucleosides, and is generally more active than morphine.

Materials and methods

Rat bone cancer model

Cell culture [0061] Cells were cultured in medium containing RPMI-1640 (Gibco, 500 ml), 10% heat inactivated fetal calf serum (Hyclone), L-glutamine (final concentration 2 mM, from company

Gibco) and antibiotic solution (final concentration 100 U / ml penicillin and 100 ug / ml streptomycin sulfate, from Gibco). Cells were released from the tissue culture flask by brief exposure to 0.1% trypsin (Gibco) and then prepared for injection as

The following occurs: the cells were centrifuged for 10 minutes at approximately 1200 rpm. The obtained granule was washed twice in phosphate buffered saline (PBS,

Mediatech) not containing calcium or magnesium. The final granule was resuspended in

PBS and cells counted using a hemocytometer. Cells were diluted to reach final concentrations for injection and kept on ice until injection.

Surgery [0062] After one week quarantine, either the culture medium or 3 x 10 was injected into the myeloid of the proximal end of the tibia.⁴ MRMT-1 rat mammary gland cancer cells. In the procedure, the animal was first anesthetized with ketamine / xylazine and the right leg area was shaved and treated with an iodine solution and cleaned with a 70% ethanol solution. A 1 cm head-caudal incision was made in the skin above the upper half of the tibia. Blunt tissue separation was performed to expose the tibia, ensuring the least damage to the muscles or blood vessels. Using a size 23 needle, the tibia was pierced 1-3 mm below the knee joint. The needle was inserted at an angle that allowed it to be pushed down into the intramedullary canal of the bone. When the access to the intramedullary canal was opened, a 23 gauge needle was removed and replaced with a blunt needle attached to a 5 μL Hamilton syringe. A volume of 3 μl culture medium + medium or tumor cells + medium was injected into the intramedullary cavity. Cancer cells were injected slowly while removing the syringe, which allowed the cells to fill the cavity. After the injection, the injection site was closed using bone wax. The wound was then closed using surgical staples. Post-surgery care and observation was carried out until the animal regained consciousness. Behavior measurements

Dosage [0063] On day 8 or 15, rats were dosed with a single injection of vehicle, reference compound or test product 20 minutes before the start of mechanical allodynia testing and approximately 40 minutes before the beginning of thermal hyperalgesia testing. Based on the pharmacological activity of SPM 927, the tests should be carried out for no longer than 90 minutes after drug administration. On day 9, rats were given a comparative compound or test article 20 minutes before the start of weight-bearing testing.

Assessment of harmful stimulus reception [0064] Pain assessment tests were performed on days 7, 8, 14 and 15. On day 7, 14 a reference value assessment was carried out. All animals received intraperitoneal injection of saline approximately 20 minutes before testing the reference value. On days 8, 15, starting approximately 20 minutes after injection of the test / comparator article, the animals underwent a series of assessments of harmful stimulus reception. The order of testing remained the same for all animals. First, mechanical allodynia was evaluated in animals, followed by thermal hyperalgesia. Weight bearing testing animals passed on days 9 and 15. First, the reference value for weight bearing response was evaluated in animals. After measuring the reference values, rats were injected with the test / comparator article and at least 20 minutes later the animals underwent another weight bearing analysis.

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Mechanical allodynia [0065] The von Frey mechanical allodynia test was performed on the affected (right) hind limb of all animals at reference point (day 7) and on day 8. In this test, rats were placed in a small Plexiglas box with a wire mesh floor. After habituating for about 10 min, a series of thin nylon fibers were applied through the floor of the cage from underneath and the rear paw plantar surface was pressed. Rats were not restrained or tethered during the test. The diameter of the filaments gave a logarithmic scale of the force exerted, and therefore a linear scale of intervals of perceived intensity. First, the weakest strength fiber was tested, which lies below the normal detection threshold for most rats. In turn, the stronger fibers were tested, in each case using the force needed only to start bending the monofilament. When the rat raised its paw in response to pressure, the filament size was recorded and then the weaker filament was used. The paw withdrawal threshold was determined according to the Chapman up-down method, involving the use of successively larger and smaller fibers to focus on the paw withdrawal threshold. A significant increase in allodynia was based on a comparison of group mean values.

Heat Hyperalgesia Test [0066] Animals underwent heat hyperalgesia testing at reference point (day 7, 14) and on day 8, 15. Each rat was placed in a separate Plexiglas chamber on an elevated heated glass surface for about 10 min for habituation. While the animal was resting, the fiber optic heat source was positioned below the glass and aimed at the right hind paw of the animal. The infrared beam was turned on, and when the rat lifted or moved its paw, the beam was turned off automatically. The stopwatch in the machine recorded a period of delay to take the foot, which was taken as meaning the time it takes the animal to detect pain due to heat. If the rat did not move within 25 seconds, the heat source automatically turned off, ensuring that the paw was not damaged. Only the affected hind paw was tested. This process was repeated at least twice for each rat, approximately 3 minutes apart. If the delays did not differ more than 2 seconds, they were averaged. If the delays differed by more than 2 seconds, the rat was tested until the two delays matched each other with an accuracy of 2 seconds and the two numbers were averaged. Group means for paw withdrawal delays were compared between groups, with a lower delay indicating greater pain sensitivity.

Weight bearing [0067] After reallocating the animals in groups 4-6 to drug treatment groups randomly, on day 9, all animals underwent weight bearing testing. Weight bearing on affected hind limbs was evaluated as the difference in weight carried by a limb on the same side compared to a limb on the opposite side. Experimentally, rats were placed in a Plexiglas chamber constructed so that each hind paw rested on a separate transducer pad that recorded the animal's body weight distribution on each paw. Five readings were taken for each of the paws, and then averaged, expressing results as a weight-bearing difference (WBD; reading on the opposite side - reading on the same side). Rats were assigned to groups of eight (8) animals per group based on body weight the day after arrival. The average body weights were checked for each group to ensure that the values

The means and standard deviations met the assumption of homogeneity. Based on the results of the von Frey test on day 8, animals in groups 4 - 6 were again randomly assigned to new treatment groups to perform a weight bearing test to prevent any deviation due to their previous assignment to groups. Animals in groups 1 - 3 remained in their designated groups, as they were animals that did not know the SPM 927 treatment.

Vincristine induced pain model

Treatment of animals [0068] 86 Dark Agouti female rats (150-200 g) were used for this study (Harlan,

Gannat, France). They were kept in groups (3 animals per cage) and kept in a temperature-controlled room (21 - 22 ° C) and with a reverse day / night cycle (12 h / 12 h) with free access to food and water. All experiments were carried out in accordance with the institution's recommendations. Vincristine poisoning was achieved by daily injection of vincristine (0.15 mg / kg / day, ip) from day 1 to 5, from day 8 to 12 and on days 15 to 16. On day 17, the animals were subjected to the behavior test and received drug treatment. Vincristine poisoned rats were randomly divided into 5 experimental groups (11 rats per group): 1. vincristine / vehicle, ip; 2. vincristine / SPM 927 (3 mg / kg), ip; 3. vincristine / SPM 927 (10 mg / kg), ip; 4. vincristine / SPM 927 (30 mg / kg), ip; 5. vincristine / morphine (3 mg / kg), sc SPM 927 and morphine were injected 30 and 45 min, respectively, before performing behavioral tests.

Cold bath test (heat allodynia) [0069] The animals were placed on an ice platform submerged approximately 1 cm below the surface of cold water (4 ° C) so that the hairy and hairless skin of the animal's foot was in contact with cold water. The period of delay before the first reaction was recorded (licking, paw movement, slight jumps), interrupting the test after 30 s.

Hot plate test (allodynia / heat hyperalgesia) [0070] The animals were placed in a glass cylinder on a hot plate (Bioblock, France) set at 38 ° C or 52 ° C. The period of delay of the first reaction (licking, pawing, slight jumps or jumping to escape from heat) was recorded, interrupting the test after 30 s. Stimulation test with von Frey filaments (mechanical allodynia) [0071] Rats were placed on a metal mesh floor. Testing for the reception of harmful stimuli was carried out by sliding the filament von Frey (Bioseb, France) through the mesh floor and applying it to the sole of the hind foot. The trial included the application of several different von Frey filaments (with a frequency of 1-1.5 s). The von Frey filaments were applied from a 10g to 100g filament. The mechanical allodynia threshold was recorded when the animal retracted its hind paw, the test was stopped and the filament number was recorded.

Paw pressure test (mechanical hyperalgesia) [0072] The bending reflex on perception of harmful stimuli was quantified using a Randall-Selitto paw pressure test device (Bioseb, France) which applies linear mechanical strength to the back of the rat's hind paw. The threshold for the value of receiving mechanical harmful stimuli was defined as the quality in grams at which the rat withdraws its paw. The final pressure was set at 250 g.

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Data analysis [0073] Analysis of variance (ANOVA) followed by post hoc analysis (Dunnett's test) was used to compare groups of behavior data at each individual time point.

Nucleoside induced pain

Animals, ddC poisoning and experimental groups [0074] 50 male Sprague Dawley rats (~ 220 g) (Janvier, Le Genest-St-Isle, France) were used for this study. The rats were housed in groups (3 animals per cage) and kept in a temperature-controlled room (21 - 22 ° C) with a reverse day / night cycle (12 h / 12 h) with free access to food and water. All experiments were carried out in accordance with the institution's recommendations. Poisoning was achieved by a single injection of ddC (50 mg / kg, iv into the tail vein). On day 10 and day 20, the animals were subjected to the behavior test and received drug treatment. DdC poisoned rats were randomly distributed into 5 experimental groups (10 rats per group): 1. control / vehicle, ip; 2. ddC / substrate, ip; 3. ddC / SPM 927 (3 mg / kg), ip; 3. ddC / SPM 927 (10 mg / kg), ip; 4. ddC / SPM 927 (30 mg / kg), ip; 5. ddC / morphine (3 mg / kg), SC SPM 927 and morphine were injected 30 and 45 min, respectively, before behavior tests.

Brushing test on day [0075] The hair on the legs, sides and bottom of the back was brushed in turn using a swab-tipped applicator using a vibrating motion (frequency 1-2 / s; 30 seconds). Brushing was done using no more than the force needed to move the applicator through the fur so as to just move the coat. Squeaks and moderate actions were counted to avoid brushing.

Results

Rat bone cancer model

Mechanical allodynia [0076] Fig. 1 shows group responses to von Frey filaments after reference value testing and after drug treatment. Statistical analysis using general two-way ANOVA was significantly different for the treatment groups (p &lt;0.01), but not for the reference value in relation to the treatment effects. One-way ANOVA tests comparing the "only cells" group after "treatment" with each treatment group revealed significant differences in mechanical allodynia levels for the morphine group (p &lt;0.01), and the 20 and 40 mg / kg SPM 927 groups (p &lt;0, 05). Treatment with 5 mg morphine / kg completely reversed allodynia as revealed during testing of the reference value. In addition, a highly significant difference was shown between the "after treatment" data for the healthy group and the "only cells" group with the injected tumor (p &lt;0.01). There are also statistical differences between the reference values ​​and post-dose values ​​for the "cell only" group and the morphine group (p &lt;0.01).

Heat hyperalgesia [0077] Data for heat tests on paws (Fig. 2) on days 14 and 15 show reference values ​​of paw withdrawal delays for all groups injected with cancer. Using two-way ANOVA, it was confirmed that this was highly significant, in which the drug's effect had p &lt;

0.0001 and the effect of treatment was significant at p &lt;0.05. Data for baseline or pre-treatment values ​​were significantly different for the "no cell" versus "only cell" group (post hoc test

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Dunnett; p &lt;0.001) with no other differences between the groups. In addition, after treatment, the groups were different for "no cells" versus "only cells" (Dunnett's test; p &lt;0.001) and "only cells" for a 30 mg / kg dose of SPM 927 (Dunnett's test; p &lt;0.001). The control group with morphine did not show statistical significance, although there was a trend indicating that morphine increased paw withdrawal delay.

Mechanical hyperalgesia [0078] The graph (Figure 3) shows bilateral ANOVA showing significant differences between groups, 0 &lt;0.001. Since the differences in weight bearing were defined as the weight transferred by the leg on the opposite side minus the weight transferred by the leg on the same side (with the tumor injected), a higher number could indicate a greater weight on the leg not affected by the disease, with 0 meaning equal weight distribution on both feet. Collected baseline data showed a positive number, revealing that all of these animals placed greater weight on their unaffected paws. Compared with the "only cells" group, after treatment the morphine group and the 40 mg / kg SPM 927 group showed a significant reduction (p &lt;0.05) of the weight placed on the paws on the opposite side compared with the "only cells" group.

Vincristine induced pain

Cold bath test [0079] As shown in Figure 4, a significant statistical difference was observed between the 6 groups (p &lt;0.05, ANOVA test). Vehicle-treated animals treated with vincristine showed a very short threshold period of delay in the cold bath test (about 9 s) as opposed to control animals showing a time result of about 14 s. Treatment of vincristine-treated animals with SPM 927 induced a significant increase (p &lt;0.05, Dunnett's test) of the delay threshold that actually became comparable to that of control animals, especially at therapeutic doses of 10 and 30 mg / kg. However, at 3 mg / kg, the threshold period of delay was slightly higher than the value for animals treated with vincristine, although no statistical difference was achieved. Treatment with morphine extended the threshold value for the delay period of vincristine-treated animals to levels well above those obtained for control rats.

Hot plate test at 38 ° C [0080] Figure 5 shows that the threshold value for the delay period of vincristine treated animals in the hot plate test (38 ° C) was significantly shorter than the value for control animals (p &lt;0.05, Dunnett test). Treatment of vincristine-treated rats with SPM 927 at doses of 3, 10 and 30 mg / kg induced a significant increase (p &lt;0.05, Dunnett's test) of the delay period threshold value. At therapeutic doses of 10 and 30 mg / kg, the performance of STZ rats became comparable to that of control animals. Like SPM 927 at 10 and 30 mg / kg, morphine at 3 mg / kg extended the STZ threshold delay period to a level comparable to that of control rats.

Hot plate test at 52 ° C [0081] As illustrated in Figure 6, the paw withdrawal delay of vincristine treated rats was significantly shorter than the delay of control animals (p &lt;0.05, Dunnett test).

Treatment of vincristine-treated rats with SPM 927 induced a significant increase (p &lt;

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0.05, Dunnett's test) paw withdrawal delay compared to vehicle treated animals. The effect obtained at doses 3 and 10, 30 mg / kg was comparable to the value for control rats.

Paw pressure test [0082] As demonstrated using the Randall &amp; Selitto anesthesia meter, vincristine treated animals showed a significant decrease in paw withdrawal delay compared to the performance of control animals (Figure 7). Treatment of vincristine-treated rats with SPM 927 at 10 and 30 mg / kg, but not at 3 mg / kg, induced a significant increase (p &lt;0.05, Dunnett's test) in paw withdrawal delay of vincristine-treated rats. In this test, morphine treatment did not modify the performance of vincristine-treated rats (p&gt; 0.05, Dunnett's test).

Von Frey filament test [0083] In this test (Figure 8), the paw withdrawal delay of vincristine treated rats was significantly reduced (about 20 g) compared to the delay for control rats (about 60 g). Treatment with SPM 927 prolonged the paw withdrawal delay of vincristine treated rats. The difference to the vehicle-treated group reached a significant level at therapeutic doses of 10 and 30 mg / kg (p &lt;0.05, Dunnett's test). Treatment with morphine restored the efficiency of vincristine-treated rats to a level comparable to the control group.

A model of rat nucleoside-induced pain

Thermal allodynia [0084] As shown in Figure 9, a significant statistical difference was observed between the 6 groups (p &lt;0.05, ANOVA test). Animals treated with ddC treated with vehicle showed a very short threshold period of delay in the cold bath test (about 11s) as opposed to control animals showing a time of about 20s. Treatment of ddC-treated animals using SPM 927 induced a significant increase (p &lt;0.05, Dunnett's test) of the delay period threshold value, which actually became comparable to the control animals at 3 test doses, 3, 10 and 30 mg / kg. Treatment with morphine extended the threshold value of the delay period of ddC-treated animals to a level comparable to that obtained with control rats.

Brushing test on day 20 [0085] Figure 10 shows the results of the brushing test carried out on day 20. Animals treated with SPM 927 (at doses of 3, 10 and 30 mg / kg) showed a significant decrease in the total number of squeaks (p &lt;0.05, Dunnett's test). Again, 3 mg / kg morphine was able to significantly reduce the number of squeaks in ddC-treated animals.

Test of von Frey filaments on day 10 [0086] In this test (Figure 11) the paw withdrawal latency of ddC-treated rats was significantly reduced (about 50 g) compared to the delay in control rats (about 85 g). Treatment with SPM 927 prolonged paw withdrawal latency in ddC rats. The difference to the vehicle treated group reached a significant level at therapeutic doses of 3, 10 and 30 mg / kg (p &lt;0.05, Dunnett's test) to a level comparable with the control group. Treatment with morphine at a dose of 3 mg / kg restored ddC-treated rats to a level similar to the control group.

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Hot plate test at 52 ° C on day 20 [0087] As illustrated in Figure 12, the paw withdrawal delay of ddC-treated rats was significantly shorter than the delay in control animals (p &lt;0.05, Dunnett test). Treatment of ddC-treated rats with SPM 927 only at 30 mg / kg induced a significant increase (p &lt;0.05, Dunnett's test) paw withdrawal latency compared to vehicle-treated animals. The effect achieved at the doses of 3 and 10 mg / kg was comparable to the untreated group to which the vehicle was administered. After morphine treatment, the performance of ddC-treated rats became similar to that of control animals.

Paw pressure test on day 10 [0088] As demonstrated with Randall &amp; Selitto anesthesia, ddC-treated animals showed a significant decrease in paw withdrawal delay compared to the performance of control animals (Figure 13). Treatment of ddC-treated rats with SPM 927 at all 3 doses of 3, 10 and 30 mg / kg induced a significant increase (p &lt;0.05, Dunnett's test) paw withdrawal latency of ddC-treated rats. Again, morphine treatment increased the efficiency of ddC-treated rats (p &lt;0.05, Dunnett's test).

Conclusion [0089] SPM 927 administered systemically gave a dose-dependent anti-allodynia and anti-hyperalgesic effect in a rat model of bone cancer pain, chemotherapy-induced pain and nucleosides, after a single dose. Thus, SPM 927 and related compounds as disclosed in formulas (Ib) or / and (IIb) are useful for treating pain during cancer, e.g. pain associated with bone cancer, after treatment with chemotherapy and nucleosides in humans.

References [0090]

Medhurst, SJ, Walker, K., Bowes, M., Kidd, B. L, Glatt, M., Muller, M., Hattenberger, M., Vaxelaire, J., O'Reilly, TO, Wotherspoon, G. , Winter, J., Green, J., and Urban, L. A rat model of bone cancer pain. Pain, 96: 129 -140, 2002.

Casey EB, Jellife AM, Le Quesne PM, Millett YL. Vincristine neuropathy. Clinical and electrophysiological observations. Brain 1973; 96: 69 - 86. Weiss HD, Walker MD, Wiernik PH. Neurotoxicity of commonly used antineoplastic agents (second of two parts). N Engl J Med.

1974; 291(3): 127 -33.

Quasthoff S, Hartung HP Chemotherapy-induced peripheral neuropathy J Neurol (2002) 249: 9 17.

Kimberly D. Tanner, David B. Reichling, and Jon D. Levine. Nociceptor Hyper-Responsiveness during Vincristine-Induced Painful Peripheral Neuropathy in the Rat. J Neurosc (1998) 18 (16): 6480 6491. Kaplan RS, Wiernik PH Neurotoxicity of antineoplastic drugs. (1982) Semin Oncol 9: 103 130.

Owellen RJ, Hartke CA, Dickerson RM, Hains FO (1976) Inhibition of tubulin-microtubule polymerization by drugs of the vinca alkaloid class. Cancer Res 36: 1499 -1502.

Sandier SG, Tobin W, Henderson ES (1969) Vincristine-induced neuropathy. A clinical study of fifty leukemic patients. Neurology 19: 367-374.

EP 1 781 276 B1

Holland JF, Scharlau C, Gailani S, Krant MJ, Olson KB, Shnider BI, Lynch JJ, Owens A, Carbone

PP, Colsky J, Grob SP, Hall TC (1973) Vincristine treatment of advanced cancer: cooperative study of 392 cases. Cancer Res 33: 1258 - 1264.

McCarthy GM, Skillings JR (1992) Jaw and other orofacial pain in patients receiving Vincristine for the treatment of cancer. Oral Surg Oral Med Oral Pathol 74: 299 -304.

Aley KO, Levine JD. Different mechanisms mediate development and expression of tolerance and dependence for peripheral mu-opioid antinociception in rat. J Neurosci 1997; 17: 8018 -23.

Aley KO, Levine JD. Role of protein kinase A in the maintenance of inflammatory pain. J Neurosci 1999; 19: 2181-6.

Aley KO, Levine JD. Different peripheral mechanisms mediate enhanced nociception in metabolic / toxic and traumatic painful peripheral neuropathies in the rat. Neuroscience 2002; 111: 389 -97.

Aley KO, McCarter G, Levine JD. Nitric oxide signaling in pain and nociceptor sensitization in the rat. J Neurosci 1998; 18: 7008 Kaplan RS, Wiernik PH Neurotoxicity of antineoplastic drugs. (1982) Semin Oncol 9: 103 -130.

Cohen J. Therapies. Confronting the limits of success. Science 2002; 296: 2320 -4.

Dalakas MC. Peripheral neuropathy and antiretroviral drugs. J Peripher Nerv Syst 2001; 6: 14-20. Dalakas MC, Semino-Mora C, Leon-Monzon M. Mitochondrial alterations with mitochondrial DNA depletion in the nerves of AIDS patients with peripheral neuropathy induced by 2030dideoxycytidine (ddC). Lab Invest 2001; 81: 1537 -44.

Dina OA, Barletta J, Chen X, Mutero A, Martin A, Messing RO, Levine JD. Key role for the epsilon isoform of protein kinase C in painful alcoholic neuropathy in the rat. J Neurosci 2000; 20: 8614 -9. Dubinsky RM, Yarchoan R, Dalakas M, Broder S. Reversible axonal neuropathy from the treatment of AIDS and related disorders with 20.30-dideoxycytidine (ddC). Muscle Nerve 1989; 12: 856 -60. Josepha EK, Chen X, Khasara SG, Levinea JD. Novel mechanism of enhanced nociception in a model of AIDS therapy-induced painful peripheral neuropathy in the rat. Pain 107 (2004) 147-158. Williams D, Geraci A, Simpson DM. AIDS and AIDS-treatment neuropathies. Curr Neural Neurosci Rep 2001; 1: 533 -8.

Yatvin MB, Li W, Meredith MJ, Shenoy MA. Improved uptake and retention of lipophilic prodrug to improve treatment of HIV. Adv Drug Deliv Rev 1999; 39: 165 -8