WO2005034873A2 - A method for reducing anti-neoplastic agent induced side-effects - Google Patents

Abstract

The present invention relates to sulfonamides, pharmaceutical compositions containing them, and their use as antagonists of urotensin II.

Description

A METHOD OF REDUCING ANTI-NEOPLASTIC AGENT INDUCED SIDE EFFECTS

FIELD OF INVENTION

The present invention relates to a method of reducing nephrotoxicity and diarrhea caused by anti-neoplastic agents in a mammal using an inhibitor (antagonist) of urotensin-II receptor.

BACKGROUND OF THE INVENTION

Effective chemotherapy for cancer treatment, which has acceptable toxicity to normal cells and tissues, is a continuing goal in the oncology field. Numerous agents are used in the treatment of cancer, including agents that adversely affect normal tissues upon use. The usefulness of virtually all anti-neoplastic drugs is severely limited by the damage they cause to these normal tissues. Several methods have been devised to reduce the side effects of anti-neoplastic agents. For example our copending patent application discloses the use of PDE4 (PDEIV) inhibitors to reduce anti-neoplastic agent induced myelosuppresion and mucositis (WO 01/93909; published December 13, 2001). Other typical side effects induced by anti-neoplastic agents, such as cisplatin (cis-diaminedichloroplatinum), include diarrhea and nephrotoxicity. Cisplatin is a heavy metal complex with a central platinum atom surrounded by two ammonia molecules and two chlorine atoms in the cis position. Cisplatin is typically used as a secondary therapy in combination with other chemotherapeutic agents for metastatic testicular tumors and metastatic ovarian tumors in the patients who have already received appropriate surgical or radiotherapeutic treatment. Cisplatin is also used as a single agent in treating patients with transitional cell bladder cancer, which is not suited for surgical, or radiotherapeutic treatment. Other uses of cisplatin include treating epithelial malignancies as well as cancers of the head and neck, the esophagus, and the lung. The major dose-limiting toxicity of cisplatin is cumulative renal insufficiency, which has been associated with renal tubular damage. Renal toxicity becomes more prolonged and more severe with repeated cisplatin treatments. Electrolyte disturbances are often secondary effects of renal damage. Hydration and diuresis are used to reduce renal toxicity, but renal damage often occurs even if these measures are taken. Urotensin-II is a vasoactive 'somatostatin-like' cyclic peptide which was originally isolated from fish spinal cords, and which has recently been cloned from man. Ames et al has identified its human receptor as hGPR14 (Ames et. al., Nature, 1999, 401, 282; Douglas & Ohlstein (2001)). Human urotensin-II is found within both vascular and cardiac tissue (including coronary atheroma) and effectively constricts isolated arteries from non-human primates. The potency of vasocontriction of urotensin-II is an order of magnitude greater than that of endothelin-1, making urotensin-II the most potent mammalian vasoconstrictor identified so far. In vivo, human urotensin-II markedly increases total peripheral resistance in anaesthetized non-human primates, a response associated with profound cardiac contractile dysfunction. Furthermore, urotensin-II immunoreactivity is also found within central nervous system and endocrine tissues. Several biological activities were proposed or found for antagonists of urotensin-II receptor (hGPR14 antagonists), see for example WO2002089792 published November 14, 2002; however until Applicants' present discovery, there has been no teaching that antagonists of urotensin-II receptor are useful in reducing side effects of nephrotoxicity and diarrhea caused by anti-neoplastic agents in a mammal.

SUMMARY OF THE INVENTION In a first aspect of the present invention, there is provided a method of reducing side effects of nephrotoxicity and/or diarrhea caused by anti-neoplastic agents in a mammal being treated with the same which comprises administering an effective amount of an urotensin-II receptor antagonist. In a second aspect of the present invention, there is provided a use of an urotensin-II receptor antagonist in the preparation of a medicament for the reduction of nephrotoxicity and/or diarrhea caused by anti-neoplastic agents in a mammal being treated with the same anti-neoplastic agents.

In a third aspect of the present invention, there is provided a pharmaceutical formulation comprising an urotensin-II receptor antagonist in an amount effective to reduce anti-neoplastic agent induced nephrotoxicity and/or diarrhea in a mammal being treated with the same anti-neoplastic agents.

DETAILED DESCRIPTION Preferred urotensin-II antagonists of the present invention are a compound of

Formula I or a pharmaceutically acceptable salt thereof

Formula I

described in our copending application WO 02/089792, published November 14, 2002, wherein:

Ri is phenyl substituted or unsubstituted by one, two , three, four or five of any of the following: halogen, CF₃, OCF₃, SCF₃, NO₂, CN,

C_1-6 alkyl, Cι.₆ alkoxy, NR₅Re, CONR₇R₈, SC_1-6 alkyl, CO₂(C₁.₆ alkyl),

C_1-6 alkyl-CO₂(C_1-6 alkyl);

R₂ is hydrogen, halogen, CF₃, CN or C alkyl;

R₃, R₄, R₇, and R₈ are independently hydrogen, C_1-6 alkyl, or benzyl;

R₅, Re, and R , are independently hydrogen or

alkyl;

X is O, S, or CH₂; n is 1.

Thus the present method relates to a method of reducing side effects of nephrotoxicity and/or diarrhea caused by anti-neoplastic agents in a mammal being treated with the same which comprises administering an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof

Formula I

wherein:

Ri is phenyl substituted or unsubstituted by one, two , three, four or five of any of the following: halogen, CF₃, OCF₃, SCF₃, NO₂, CN, Cι-6 alkyl, C_1-6 alkoxy, NR₅R<., CONR₇Rs, SC_1-6 alkyl, CO₂(C_1-6 alkyl),

Cι-₆ alkyl-CO₂(C_1-6 alkyl);

R₂ is hydrogen, halogen, CF₃, CN or C_1- alkyl;

R₃, t, R , and R₈ are independently hydrogen, C_1-6 alkyl, or benzyl;

R₅, R_ό, and R₉, are independently hydrogen or Cι_₆ alkyl; X is O, S, or CH₂; and n is 1.

Furthermore, the present invention relates to a pharmaceutical formulation comprising a compound of Formula I or a pharmaceutically acceptable salt thereof

Formula I

wherein: Ri is phenyl substituted or unsubstituted by one, two , three, four or five of any of the following: halogen, CF₃, OCF₃, SCF₃, NO₂, CN, Cι-₆ alkyl, C_1-6 alkoxy, NR₅Re, CONR₇Rs, SC_1-6 alkyl, CO₂(C_1-6 alkyl), Cι-₆ alkyl-CO₂(C₁.₆ alkyl); R₂ is hydrogen, halogen, CF₃, CN or Cμ alkyl;

R₃, R₄, R , and R₈ are independently hydrogen, C_1-6 alkyl, or benzyl; R₅, Re_. and R , are independently hydrogen or Cι.₆ alkyl; X is O, S, or CH₂; and n is 1; in an amount effective to reduce anti-neoplastic agent induced nephrotoxicity and/or diarrhea in a mammal being treated with the same anti-neoplastic agent.

Even more preferred urotensin-II receptor antagonists are a compound of

Formula I in which Ri is phenyl substituted or unsubstituted by one, two , three, four, or five of any of the following: halogen, CF₃, OCF₃, CN,

Cι-6 alkyl, C₁.₆ alkoxy, CO₂(C_1-6 alkyl), C_1-6 alkyl-CO₂(C_1-6 alkyl), orNO₂;

R₂ is hydrogen, halogen, CF₃, or C_1-4 alkyl;

R₃ is hydrogen; R₄ is hydrogen;

R is hydrogen or C_1-6 alkyl; and

X is O.

Yet even more preferred urotensin-II receptor antagonists of the instant invention are (R2-Bromo-N-[4-chloro-3-((R)-l-methyl-pyrrolidin-3-yloxy)-phenyl]- 4,5-dimethoxy-benzenesulfonamide of Formula la

la

and 2-bromo-4,5-dimethoxy-N-[3-(l-methyl-pyrrolidin-3-yloxy-4-trifluormethy- phenyl]-benzenesulfonamide of Formula lb

lb

Thus, there is provided a pharmaceutical formulation comprising a compound of Formula la or lb, or a pharmaceutically acceptable salt thereof, in an amount effective to reduce anti-neoplastic agent induced nephrotoxicity and/or diarrhea in a mammal being treated with the same anti-neoplastic agents. In another aspect, there is provided a use of a compound of Formula I, la or lb in the preparation of a medicament for the reduction of nephrotoxicity and/or diarrhea caused by anti-neoplastic agents in a mammal being treated with the same anti- neoplastic agents.

When used herein, the term "alkyl" includes all straight chain and branched isomers. Representative examples thereof include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, t-butyl, n-pentyl and n-hexyl. When used herein, the terms 'halogen' and 'halo' include fluorine, chlorine, bromine and iodine, and fluoro, chloro, bromo and iodo, respectively. The compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active form. All of these compounds and their diastereoisomers are contemplated to be within the scope of the present invention. The syntheses of compounds of Formula I, la or lb are all well described in WO

02/089792. As used herein "urotensin-II" refers to human eleven amino acid peptide with amino acid sequence ETPDCFWKYCV (SEQ ID NO:l). Urotensin-II receptor refers to 7-transmembrane receptor which is also referred to as hGPR14 (GenBank accession number AF140631) reported in Nature, 1999, 401,

282; Douglas & Ohlstein (2001). Many urotensin-II receptor antagonists (or hGPR14 antagonists) have already been reported and are well known. The following patent publications discloses examples of urotensin-II receptor antagonists:

WO2002076979, published October 3, 2002

WO200166143; published March 6, 2001;

WO200202530, published January 10, 2002;

WO200215934, published February 28, 2002; WO200232932, published April 25, 2002;

WO2002090353, published November 14, 2002

WO2002090348, published November 14, 2002

WO2002089793, published November 14, 2002

WO2002089792, published November 14, 2002 WO2002089740, published November 14, 2002

WO2002089740, published November 14, 2002

WO2002090337, published November 14, 2002:

WO2002078641, published November 14, 2002

WO2002079155, published October 10, 2002; WO2002079188, published October 10, 2002;

WO2002078707, published October 10, 2002;

WO2002058702, published August 1, 2002; WO200247456, published June 20, 2002; WO200247687, published June 20, 2002; WO200145694, published June 28, 2001; WO200245700, published June 28, 2001; WO200145711, published June 28, 2001; WO200137780, published May 31, 2001; and WO200137856, published May 31 , 2001.

As used herein urotensin-II receptor antagonists of the present invention include, but are not limited to, any of the compounds disclosed in the above patent publications.

As used herein the term "neoplasm" refers to an abnormal growth of cells or tissue and is understood to include benign, i.e., non-cancerous growths, and malignant, i.e., cancerous growths. The term "neoplastic" means of or related to a neoplasm. As used herein the term "agent" is understood to mean a substance that produces a desired effect in a tissue, system, animal, mammal (in particular human), or other subject. Accordingly, the term "anti-neoplastic agent" is understood to mean a substance producing an anti-neoplastic effect in a tissue, system, animal, mammal (in particular human), or other subject. It is also to be understood that an "agent" may be a single compound or a combination or composition of two or more compounds. Some of the typical anti-neoplastic agents include alkylating agents such as melphalan, chlorambucil, cyclophosphamide, mechlorethamine, hexamethylmelamine, busulfan, carmustine, lomustine, and dacarbazine; antimetabolites such as 5- fluorouracil, methotrexate, cytarabine, mecaptopurine and thioguanine; antimitotic agents such as paclitaxel, docetaxel, vinblastine, vincristine; topoisomerase I inhibitors such as irinotecan, camptothecin and camptothecin derivatives; topoisomerase II inhibitors such as doxorubicin; and platinum coordination complexes such as cisplatin and carboplatin. As used herein, the term "effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal, mammal (including human), or other subject that is being sought by a researcher or clinician. Furthermore the term "therapeutically effective amount" means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder, and also includes amounts effective to enhance normal physiological function. More specifically typical anti-neoplastic agents for which the present urotensin-

II receptor antagonists are useful for reducing nephrotoxicity and diahrrea include, cisplatin, diterpenoids, vinca alkaloids, epipodophyllotoxins, antimetabolites, and camptothecins. Diterpenoids, which are derived from natural sources, are phase specific anti - cancer agents that operate at the G2/M phases of the cell cycle. It is believed that the diterpenoids stabilize the β-tubulin subunit of the microtubules, by binding with this protein. Disassembly of the protein appears then to be inhibited with mitosis being arrested and cell death following. Examples of diterpenoids include, but are not limited to, paclitaxel and its analog docetaxel. Paclitaxel, 5β,20-epoxy-l,2α,4,7β,10β,13α-hexa-hydroxytax-l l-en-9-one 4,10- diacetate 2-benzoate 13-ester with (2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. It is a member of the taxane family of terpenes. It was first isolated in 1971 by Wani et al. J. Am. Chem, Soc, 93:2325. 1971), who characterized its structure by chemical and X-ray crystallographic methods. One mechanism for its activity relates to paclitaxel's capacity to bind tubulin, thereby inhibiting cancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA, 77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar, J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis and anticancer activity of some paclitaxel derivatives see: D. G. I. Kingston et al., Studies in Organic Chemistry vol. 26, entitled "New trends in Natural Products Chemistry 1986", Attaur-Rahman, P.W. Le Quesne, Eds. (Elsevier, Amsterdam, 1986) pp 219- 235. Paclitaxel has been approved for clinical use in the treatment of refractory ovarian cancer in the United States (Markman et al., Yale Journal of Biology and Medicine, 64:583, 1991; McGuire et al, Ann. Intern, Med., 111:273,1989) and for the treatment of breast cancer (Holmes et al, J. Nat. Cancer Inst, 83:1797,1991.) It is a potential candidate for treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al, Sein. Oncol., 20:56, 1990). The compound also shows potential for the treatment of polycystic kidney disease (Woo et al., Nature, 368:750. 1994), lung cancer and malaria. Docetaxel, (2R,3S)- N-carboxy-3-phenylisoserine,N-tert-butyl ester, 13-ester with 5β-20-epoxy-l,2α,4,7β,10β,13α-hexahydroxytax-ll-en-9-one 4-acetate 2- benzoate, trihydrate; is commercially available as an injectable solution as TAXOTERE®. Docetaxel is indicated for the treatment of breast cancer. Docetaxel is a semisynthetic derivative of paclitaxel q.v., prepared using a natural precursor, 10- deacetyl-baccatin III, extracted from the needle of the European Yew tree. Vinca alkaloids are phase specific anti-neoplastic agents derived -from the periwinkle plant. Vinca alkaloids act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, and vinorelbine. Vinblastine, vincaleukoblastine sulfate, is commercially available as

VELBAN® as an injectable solution. Although, it has possible indication as a second line therapy of various solid tumors, it is primarily indicated in the treatment of testicular cancer and various lymphomas including Hodgkin's Disease; lymphocytic and histiocytic lymphomas. Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commercially available as ONCOVIN® as an injectable solution. Vincristine is indicated for the treatment of acute leukemias and has also found use in treatment regimens for Hodgkin's and non- Hodgkin's malignant lymphomas. Vinorelbine, 3',4'-didehydro -4'-deoxy-C'-norvincaleukoblastine [R-(R*,R*)- 2,3-dihydroxybutanedioate (l:2)(salt)], commercially available as an injectable solution of vinorelbine tartrate (NAVELBINE®), is a semisynthetic vinca alkaloid. Vinorelbine is indicated as a single agent or in combination with other chemotherapeutic agents, such as cisplatin, in the treatment of various solid tumours, particularly non-s nall cell lung, advanced breast, and hormone refractory prostate cancers. Epipodophyllotoxins are phase specific anti-neoplastic agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing

DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide and teniposide. Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R )-ethylidene-β-D- glucopyranoside], is commercially available as an injectable solution or capsules as

VePESID® and is commonly known as VP-16. Etoposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of testicular and non-small cell lung cancers. Teniposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R. )-thenylidene-β-D- glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26. Teniposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia in children. Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows. Examples of antimetabolite anti-neoplastic agents include, but are not limited to, fluorouracil, methotrexate, cytarabine, mecaptopurine and thioguanine. 5-Fluorouracil, 5-fluoro-2,4- (1H,3H) pyrimidinedione, is commercially available as fluorouracil. Adminsitration of 5-fluorouracil leads to inhibition of thymidylate synthesis and is also incorporated into both RNA and DNA. The result typically is cell death. 5-Fluorouracil is indicated as a single agent or in combination with other chemotherapy agents in the treatment of carcinomas of the breast, colon, rectum, stomach and pancreas. Other fluoropyrimidine analogs include 5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridine monophosphate. Cytarabine, 4-amino-l-β-D-arabinofuranosyl-2 (lH)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. It is believed that cytarabine exhibits cell phase specificity at S-phase by inhibiting DNA chain elongation by terminal incorporation of cytarabine into the growing DNA chain. Cytarabine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Other cytidine analogs include 5-azacytidine and 2',2'-difluorodeoxycytidine (gemcitabine). Mercaptopurine, l,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®. Mercaptopurine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Mercaptopurine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppresion and gastrointestinal mucositis are expected side effects of mercaptopurine at high doses. A useful mercaptopurine analog is azathioprine. TWoguanine, 2-amino-l,7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®. Thioguanine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Thioguanine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Other purine analogs include pentostatin, erythrohydroxynonyladenine, fludarabine phosphate, and cladribine. Methotrexate, N-[4[[(2,4-diamino-6-pteridinyl) methyljmethylamino] benzoyl]- L-glutamic acid, is commercially available as methotrexate sodium. Methotrexate exhibits cell phase effects specifically at S-phase by inhibiting DNA synthesis, repair and/or replication through the inhibition of dyhydrofolic acid reductase which is required for synthesis of purine nucleotides and thymidylate. Methotrexate is indicated as a single agent or in combination with other chemotherapy agents in the treatment of choriocarconoma, meningeal leukemia, non-Hodgkin's lymphoma, and carcinomas of the breast, head, neck, ovary and bladder. Camptothecins, including, camptothecin and camptothecin derivatives are under development as Topoisomerase I inhibitors. Camptothecins cytotoxic activity is believed to be related to its Topoisomerase I inhibitory activity. Examples of camptothecins include, but are not limited to irinotecan and the various optical forms of 7-(4-methylpiperazino-methylene)- 10, 11 -ethylenedioxy-20-camptothecin. Irinotecan HC1, (4S)-4, 11 -diethyl-4-hydroxy-9-[(4-piperidinopiperidino) carbonyloxy]-lH-pyrano[3',4',6,7]indolizino[l,2-b]quinoline-3,14(4H,12H)-dione hydrochloride, is commercially available as the injectable solution CAMPTOSAR®. Irinotecan is a derivative of camptothecin which binds, along with its active metabolite SN-38, to the topoisomerase I - DNA complex. It is believed that cytotoxicity occurs as a result of irreparable double strand breaks caused by interaction of the topoisomerase I : DNA : irintecan or SN-38 ternary complex with replication enzymes. Irinotecan is indicated for treatment of metastatic cancer of the colon or rectum. The dose limiting side effects of irinotecan HC1 are myelosuppression, including neutropenia, and GI effects, including diarrhea.

The present use of urotensin-II receptor antagonists to reduce anti-neoplastic agents induced sides effects are for both human and veterinary use. However, it is well understood that the effectiveness of a particular urotensin-II receptor antagonist in a particular species is limited, such as, by its bioavailability in that species. Which urotensin-II receptor antagonist is more effective in a particular species can be readily ascertained by conventional pharmacological methods.

The anti-neoplastic agent and the urotensin-II receptor antagonist may be employed in combination in accordance with the invention by administration concomitantly in (1) a unitary pharmaceutical composition including both compounds or (2) separate pharmaceutical compositions each including one of the compounds. Alternatively, the combination may be administered separately in a sequential manner wherein the urotensin-II receptor antagonist or anti-neoplastic agent is administered first and the other second. Such sequential administration may be close in time or remote in time. In a presently preferred embodiment, the urotensin-II receptor antagonist is administered first and the anti-neoplastic agent is subsequently administered within a time period suitable to achieve the desired therapeutic effect. The present method of using urotensin-II receptor antagonists to reduce anti-neoplastic agent induced diarrhea and nephrotoxicity is applicable in any situations when anti-neoplastic agents are being administered to treat cancers or tumors. However, the most preferred situation is when tumors or cancers being treated are those of solid malignancies, notably those of the bladder, cervix, lung, ovary, and testis such as testicular tumor; bladder cancer; ureterpyelonephritic tumor; prostatic cancer; ovarian cancer; head and neck cancer; non-small-cell lung cancer; esophageal cancer; cervical cancer; neuroblastoma; gastric cancer; small cell lung cancer; bone cancer; non-Hodgkin's lymphomas; tumours of brain, endometrium, upper gastrointestinal tract, head and neck, and thymus; neuroblastoma; and sarcoma of bone and soft tissue. The urotensin-II receptor antagonist of the present invention may be administered by any appropriate route. Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), vaginal, and parenteral (including subcutaneous, intramuscular, intraveneous, intradermal, intrathecal, and epidural). It will be appreciated that the preferred route may vary with, for example, the condition of the recipient. Pharmaceutical formulations adapted for oral administration may be presented as discrete units such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or whips; or oil-in-water liquid emulsions or water-in-oil liquid emulsions. For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with, an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Powders are prepared by comminuting the compound to a suitable fine size and mixing with a similarly comminuted pharmaceutical carrier such as an edible carbohydrate, as, for example, starch or mannitol. Flavoring, preservative, dispersing and coloring agent can also be present. Capsules are made by preparing a powder mixture as described above, and filling formed gelatin sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate or solid polyethylene glycol can be added to the powder mixture before the filling operation. A disintegrating or solubilizing agent such as agar-agar, calcium carbonate or sodium carbonate can also be added to improve the availability of the medicament when the capsule is ingested. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Tablets are formulated, for example, by preparing a powder mixture, granulating or slugging, adding a lubricant and disintegrant and pressing into tablets. A powder mixture is prepared by mixing the compound, suitably comminuted, with a diluent or base as described above, and optionally, with a binder such as carboxymethylcellulose, an aliginate, gelatin, or polyvinyl pyrrolidone, a solution retardant such as paraffin, a resorption accelerator such as a quaternary salt and/or an absorption agent such as bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acadia mucilage or solutions of cellulosic or polymeric materials and forcing through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and the result is imperfectly formed slugs broken into granules. The granules can be lubricated to prevent sticking to the tablet forming dies by means of the addition of stearic acid, a stearate salt, talc or mineral oil. The lubricated mixture is then compressed into tablets. The compounds of the present invention can also be combined with free flowing inert carrier and compressed into tablets directly without going through the granulating or slugging steps. A clear or opaque protective coating consisting of a sealing coat of shellac, a coating of sugar or polymeric material and a polish coating of wax can be provided. Dyestuffs can be added to these coatings to distinguish different unit dosages. Oral fluids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared by dissolving the compound in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added. Where appropriate, dosage unit formulations for oral administration can be microencapsulated. The formulation can also be prepared to prolong or sustain the release as for example by coating or embedding particulate material in polymers, wax or the like. The agents for use according to the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. Pharmaceutical formulations adapted for transdermal administration may be presented as discrete patches intended to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. For example, the active ingredient may be delivered from the patch by iontophoresis as generally described in Pharmaceutical

Research, 3(6), 318 (1986). Pharmaceutical formulations adapted for rectal administration may be presented as suppositories or as enemas. Pharmaceutical formulations adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets. It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents. Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting a free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base. Also, contemplated in the present invention is a pharmaceutical combination including an anti-neoplastic agent and an urotensin-II receptor antagonist. Typically, the therapeutically effective amount of an urotensin-II receptor antagonist of the present invention will depend upon a number of factors including, for example, the age and weight of the mammal, the precise condition requiring treatment, the severity of the condition, the nature of the formulation, and the route of administration. Ultimately, the therapeutically effective amount will be at the discretion of the attendant physician or veterinarian. Typically, the anti-neoplastic agent will be given in the range of 0.1 to 100 mg kg body weight of recipient (mammal) per day and more usually in the range of 1 to 30 mg/kg body weight per day. Acceptable daily dosages of the urotenin-II receptor antagonist for preventing/reducing the severity of side effects induced by administration of an anti-neoplastic agent, may be from about 0.1 to about 1000 mg/day, and preferably from about 0.2 to about 100 mg/day. The following examples are intended for illustration only and are not intended to limit the scope of the invention in any way. The following abbreviations are used in the examples following as well as in other portions of the specification.

BUNT blood urea nitrogen LDH lactate dehydrogenase γ-GTP γ-guanocin triphosphate

Example 1

Preparation of Compound A (Formula lb compound hydrochloride salt) Suspension. Compound A was suspended in 1% MC-Na/water solution with a sonicator and utilized in the following examples. Cisplatin-induced nephrotoxicity was assessed by both rise in serum creatinine and BUN levels that reflect renal dysfunction and increased excretion of urinary LDH and γ-GTP that exhibit renal tissue damage. Prevention of cisplatin-induced nephrotoxicity was shown by suppression of such increase in those serum and urinary parameters. Prevention of diarrhea caused by cisplatin treatment was inhibition of diarrlieal fecal condition. Intraperitoneal treatment of the mice with 15 mg/kg of cisplatin induced rises in serum creatinine and BUN levels and urinary LDH and γ-GTP excretion. At the same time, cisplatin treatment also caused diarrhea. In Examples 2 and 3, Compound A was tested for its ability to inhibit cisplatin-induced nephrotoxicity and diarrhea, respectively.

Example 2

Effect of Compound A on cisplatin-induced nephrotoxicity in mice. Method: Male BALB/c mice, 5 weeks of age, were used for the experiment. Compound A suspended in 1% MC-Na/water (vehicle) was orally administered to the mice at the doses of 10-100 mg/kg 1 hour prior to the cisplatin treatment. The control group was administered with the vehicle. Then, the mice were intraperitoneally treated with cisplatin at 15 mg/kg (Day 0). The urine was collected on Day 3 for analysis of urinary parameters. On Day 4, the mice were bled from the abdominal vein under anesthesia for analysis of serum parameters. Urinary LDH, γ-GTP and creatinine levels and the concentrations of serum BUN and creatinine were determined with a biochemical auto-analyzer for assessment of the tissue damage and dysfunction of kidney. All the groups consisted of 6 animals (Ishikawa, M. et al., (1993), Biol. Farm. Bull 16:1104-1107). Oral treatment of the mice with Compound A dose-dependently prevented cisplatin-induced nephrotoxicity (Table 1).

Table 1. Effects of Compound A on cisplatin-induced nephrotoxicity in mice. Dose Route N Serum Serum BUN Urinary LDH Urinary y -GTP (mg/ Creatinine (mmol L) (IU/g creatinine) (IU/g creatinine) kg) ( μ mol/L) Intact - - 8 9.23+0.39 8.30+0.30 134.30+9.71 277.01 ±15.61 Vehicle - p.o. 8 S9.37± 100.72+ 1980.55+: 2898.52+ 10.74## 17.82## 240.41## 226.52## Compound A 10 p.o. 7-8 62.21 ± 103.28+12.75 2088.63+226.80 2837.41 + 159 11.29 Compound A 30 p.o. 27.04+ 35.81 + 1318.55±181.67 1817.78+ 3.33* 10.35** 234.21** Compound A 100 p.o. 7-8 20.36+ 14.23 + 1.85** 1168.02+ 1818.51 + 1.18** 132.34* 254.89**

Each data represents the mean ± SEM (n=7~ 8). ##p<0.01: compared with Intact (Student's t-test).

*p<0.05, **p<0.01: compared with Vehicle (Dunnett's multiple comparison test).

Example 3

Effect of Compound A on cisplatin-induced diarrhea in mice. Method: Male BALB/c mice, 5 weeks of age, were used for the experiment. Compound A suspended in 1% MC-Na/water (vehicle) was orally administered to the mice at at the doses of 10-100 mg/kg 1 hour prior to the cisplatin treatment. The control group was administered with the vehicle. Then, the mice were intraperitoneally treated with cisplatin at 15 mg/kg (Day 0). On Day 4, diarrheal symptoms were scored (incidence and rating: 0: normal, 1: wet but formed stools (slight diarrhea), and 2: swollen and or unformed stools (diarrhea)). All the groups consisted of 6 animals (Ishikawa, M. et al., (1993), Biol. Pharm. Bull 16:1104-1107). Oral treatment of the mice with Compound A dose-dependently prevented cisplatin-induced diarrhea (Table 2).

Table 2. Effects of Compound A on cisplatin-induced diarrhea in mice. Dose Route n Diarrheal Diarrheal (mg/kg) score incidence Intact - - 8 0.0 0/8 Vehicle - p.o. 8 2.0## 8/8 Compound A 10 p.o. 8 2.0 8/8 Compound A 30 p.o. 8 1.6 7/8 Compound A 100 p.o. 8 1.4* 7/8

##p<0.01 : compared with Intact (Mann- Whitney U-test).

*p<0.05 : compared with Vehicle (Non-parametric Dunnett's multiple

Claims

What is claimed is:

1. A method of reducing side effects of nephrotoxicity and/or diarrhea caused by anti- neoplastic agents in a mammal being treated with the same which comprises administering an effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof

Formula I

wherein:

Ri is phenyl substituted or unsubstituted by one, two , three, four or five of any of the following: halogen, CF₃, OCF₃, SCF₃, NO₂, CN,

Ct-₆ alkyl, C,.₆ alkoxy, NRsRe, CONR₇R₈, SC_1-6 alkyl, CO₂(C_1-6 alkyl),

C_1-6 alkyl-CO₂(Ct.₆ alkyl);

R₂ is hydrogen, halogen, CF₃, CN or C_1- alkyl;

R₃, R , R₇, and R₈ are independently hydrogen, Cι.₆ alkyl, or benzyl;

R₅, Re, and R₉, are independently hydrogen or C_1-6 alkyl;

X is O, S, or CH₂; and n is 1.

2. A pharmaceutical formulation comprising a compound of Formula I or a pharmaceutically acceptable salt thereof

Formula I wherein:

Ri is phenyl substituted or unsubstituted by one, two , three, four or five of any of the following: halogen, CF₃, OCF₃, SCF₃, NO₂, CN, Cι-6 alkyl, C_1-6 alkoxy, NR₅Re, CONR₇R8, SC_1-6 alkyl, CO₂(C_1-6 alkyl), C_1-6 alkyl-CO₂(C_1-6 alkyl); R₂ is hydrogen, halogen, CF₃, CN or C alkyl; R₃, R₄, R₇, and R₈ are independently hydrogen, C_1-6 alkyl, or benzyl; R₅, Re, and R , are independently hydrogen or C1-6 alkyl; X is O, S, or CH₂; and n is 1; in an amount effective to reduce anti-neoplastic agent induced nephrotoxicity and/or diarrhea in a mammal being treated with the same anti-neoplastic agent.

3. A compound of Formula I in claim 1 or 2 in which

Ri is phenyl substituted or unsubstituted by one, two , three, four, or five of any of the following: halogen, CF₃, OCF₃, CN,

C_1-6 alkyl, C_1-6 alkoxy, CO₂(C_1-6 alkyl), C_1-6 alkyl-CO₂(C_1-6 alkyl), orNO₂;

R₂ is hydrogen, halogen, CF₃, or C_M alkyl; R₃ is hydrogen;

R₄ is hydrogen;

R₉ is hydrogen or C_1-6 alkyl; and

X is O.

4. The compound of claim 3 that is a compound of Formula la

la

5. The compound of claim 3 that is a compound of Formula lb

lb