A method for inhibiting neoplasia, particularly cancerous and precancerous lesions by exposing the affected cells to isoquinoline derivatives.

This invention relates to a method for the selective inhibition of neoplastic cells, for example, for the treatment or prevention of precancerous lesions or other neoplasias in mammals.
Each year in the United States alone, untold numbers of people develop precancerous lesions, which is a form of neoplasia, as discussed below. Such lesions exhibit a strong tendency to develop into malignant tumors, or cancer. Such lesions include lesions of the breast (that can develop into breast cancer), lesions of the skin (that can develop into malignant melanoma or basal cell carcinoma), colonic adenomatous polyps (that can develop into colon cancer), and other such neoplasms. Compounds that prevent or induce the remission of existing precancerous or cancerous lesions or carcinomas would greatly reduce illness and death from cancer.
For example, approximately 60,000 people die from colon cancer, and over 150,000 new cases of colon cancer are diagnosed each year. For the American population as a whole, individuals have a six percent lifetime risk of developing colon cancer, making it the second most prevalent form of cancer in the country. Colon cancer is also prevalent in Western Europe. It is believed that increased dietary fat consumption is increasing the risk of colon cancer in Japan.
In addition, the incidence of colon cancer reportedly increases with age, particularly after the age of 40. Since the mean ages of populations in America and Western Europe are increasing, the prevalence of colorectal cancer should increase in the future.
To date, little progress has been made in the prevention and treatment of colorectal cancer, as reflected by the lack of change in the five-year survival rate over the last few decades. The only cure for this cancer is surgery at an extremely early stage. Unfortunately, most of these cancers are discovered too late for surgical cure. In many cases, the patient does not experience symptoms until the cancer has progressed to a malignant stage.
In view of these grim statistics, efforts in recent years have concentrated on colon cancer prevention. Colon cancer usually arises from pre-existing benign neoplastic growths known as polyps. Prevention efforts have emphasized the identification and removal of colonic polyps. Polyps are identified by x-ray and/or colonoscopy, and usually removed by devices associated with the colonoscope. The increased use of colon x-rays and colonoscopies in recent years has detected clinically significant precancerous polyps in four to six times the number of individuals per year that acquire colon cancer. During the past five years alone, an estimated 3.5 to 5.5 million people in the United States have been diagnosed with adenomatous colonic polyps, and it is estimated that many more people have or are susceptible to developing this condition, but are as yet undiagnosed. In fact, there are estimates that 10-12 percent of people over the age of 40 will form clinically significant adenomatous polyps.
Removal of polyps has been accomplished either with surgery or fiber-optic endoscopic polypectomyxe2x80x94procedures that are uncomfortable, costly (the cost of a single polypectomy ranges between $1,000 and $1,500 for endoscopic treatment and more for surgery), and involve a small but significant risk of colon perforation. Overall, about $2.5 billion is spent annually in the United States in colon cancer treatment and prevention.
In the breast, breast cancer is often treated surgically, often by radical mastectomy with its painful aftermath Such surgery is costly, too.
As indicated above, each lesion carries with it a chance that it will develop into a cancer. The likelihood of cancer is diminished if a precancerous lesion is removed. However, many of these patients demonstrate a propensity for developing additional lesions in the future. They must, therefore, be monitored periodically for the rest of their lives for reoccurrence.
In most cases (i.e. the cases of sporadic lesion formation, e.g. so-called common sporadic polyps), lesion removal will be effective to reduce the risk of cancer. In a small percentage of cases (i.e. cases where numerous lesions form, e.g. the so-called polyposis syndromes), removal of all or part of the effected area (e.g. the colon) is indicated. For example, the difference between common sporadic polyps and polyposis syndromes is dramatic. Common sporadic polyp cases are characterized by relatively few polyps which can usually be removed leaving the colon intact. By contrast, polyposis syndrome cases can be characterized by many (e.g. hundreds or more) of polypsxe2x80x94literally covering the colon in some casesxe2x80x94making safe removal of the polyps impossible short of surgical removal of the colon.
Because each lesion carries with it a palpable risk of cancerous development, patients who form many lesions (e.g. polyposis syndrome patients) invariably develop cancer if left untreated. Surgical removal of the colon is the conventional treatment in polyposis patients. Many polyposis patients have undergone a severe change in lifestyle as a result of the disfiguring surgery. Patients have strict dietary restrictions, and many must wear ostomy appliances to collect their intestinal wastes.
The search for drugs useful for treating and preventing cancer is intensive. Indeed, much of the focus of cancer research today is on the prevention of cancer because chemotherapy for cancer itself is often not effective and has severe side effects. Cancer chemoprevention is important for recovered cancer patients who retain a risk of cancer reoccurrence. Also, cancer prevention is important for people who have not yet had cancer, but have hereditary factors that place them at risk of developing cancer. With the development of new genetic screening technologies, it is easier to identify those patients with high-risk genetic factors, such as the potential for polyposis syndrome, who would greatly benefit from chemopreventative drugs. Therefore, finding such anti-cancer drugs that can be used for prolonged preventive use is of vital interest.
Known chemopreventative and chemotherapeutic drugs are believed to kill cancer cells by inducing apoptosis, or as sometimes referred to as xe2x80x9cprogrammed cell death.xe2x80x9d Apoptosis naturally occurs in virtually all tissues of the body, and especially in self-renewing tissues such as bone marrow, immune cells, gut, liver and skin. Apoptosis plays a critical role in tissue homeostasis, that is, it ensures that the number of new cells produced are correspondingly offset by an equal number of cells that die. For example, the cells in the intestinal lining divide so rapidly that the body must eliminate cells after only three days in order to prevent the overgrowth of the intestinal lining.
Recently, scientists have realized that abnormalities of apoptosis can lead to the formation of precancerous lesions and carcinomas. Also, recent research indicates that defects in apoptosis play a major role in other diseases in addition to cancer. Consequently, compounds that modulate apoptosis could be used to prevent or control cancer, as well as used in the treatment of other diseases.
Unfortunately, even though known chemotherapeutic drugs may exhibit such desirable apoptosis effects, most chemotherapeutic drugs have serious side effects that prohibit their long-term use, or use in otherwise healthy individuals with precancerous lesions, These side effects, which are a result of the high levels of cytotoxicity of the drugs, include hair loss, weight loss, vomiting, immune suppression and other toxicities. Therefore, there is a need to identify new drug candidates for therapy that do not have such serious side effects in humans.
In recent years, several non-steroidal anti-inflammatory drugs (xe2x80x9cNSAIDsxe2x80x9d), originally developed to treat arthritis, have shown effectiveness in inhibiting and eliminating colonic polyps. Polyps virtually disappear when the patients take the drug, particularly when the NSAID sulindac is administered. However, the prophylactic use of currently available NSAIDs, even in polyposis syndrome patients, is marked by severe side reactions that include gastrointestinal irritations, perforations, ulcerations and kidney toxicity. Once NSAID treatment is terminated due to such complications, the polyps return, particularly in polyposis syndrome patients.
Sulindac has been particularly well received among the NSAIDs for the polyp treatment. Sulindac is a sulfoxide compound that itself is believed to be inactive as an anti-arthritic agent. The sulfoxide is reportedly converted by liver enzymes to the corresponding sulfide, which is acknowledged to be the active moiety as a prostaglandin synthesis inhibitor. The sulfide, however, is associated with the side effects of conventional NSAIDs. The sulfoxide is also known to be metabolized to sulfone compound that has been found to be inactive as an inhibitor of prostaglandin synthesis but active as an inhibitor of precancerous lesions.
This invention includes a method of inhibiting neoplastic cells by exposing those cells to a pharmacologically effective amount of those compounds described below. Such compounds are effective in modulating apoptosis and eliminating and inhibiting the growth of neoplasias such as precancerous lesions, but are not characterized by the severe side reactions of conventional NSAIDs or other chemotherapeutics.
The compounds of that are useful in the methods of this invention include those of Formula I:
wherein
Ring A and Ring B are the same or different and each a substituted or unsubstituted benzene ring, R1 is (1) a hydrogen atom, (2) a substituted or unsubstituted lower alkyl group, (3) a substituted or unsubstituted cyclo-lower alkyl group, (4) a substituted or unsubstituted aryl group, (5) a substituted or unsubstituted heterocyclic group, or (6) an amino group optionally having one or two substituents, R2 is a group of the formula xe2x80x94COOR3 or xe2x80x94CON(R4)(R5), R3 is a hydrogen atom or an ester residue, and a group of the formula xe2x80x94N(R4)(R5) is a substituted or unsubstituted nitrogen-containing aliphatic heterocyclic group or a substituted or unsubstituted amino group, provided that when R1 is a hydrogen atom or a substituted or unsubstituted lower alkyl group, then at least one of Ring A and Ring B is a benzene ring which is substituted by two or more lower alkoxy groups, or a pharmaceutically acceptable salt thereof.
As indicated above, this invention relates to a method for inhibiting neoplasia, particularly cancerous and precancerous lesions by exposing the affected cells to a compound of Formula I above.
Preferably, such compounds are administered without therapeutic amounts of an NSAID.
The present invention is also a method of treating mammals with precancerous lesions by administering a pharmacologically effective amount of an enterically coated pharmaceutical composition that includes compounds of this invention.
Also, the present invention is a method of inhibiting the growth of neoplastic cells by exposing the cells to an effective amount of compounds of Formula I, wherein R1 through R3 are defined as above.
In still another form, the invention is a method of inducing apoptosis in human cells by exposing those cells to an effective amount of compounds of Formula I to those cells sensitive to such a compound.
As used herein, the term xe2x80x9cprecancerous lesionxe2x80x9d includes syndromes represented by abnormal neoplastic, including dysplastic, changes of tissue.
Examples include adenomatous growths in colonic, breast or lung tissues, or conditions such as dysplastic nevus syndrome, a precursor to malignant melanoma of the skin. Examples also include, in addition to dysplastic nevus syndromes, polyposis syndromes, colonic polyps, precancerous lesions of the cervix (i.e., cervical dysplasia), prostatic dysplasia, bronchial dysplasia, breast, bladder and/or skin and related conditions (e.g., actinic keratosis), whether the lesions are clinically identifiable or not.
As used herein, the term xe2x80x9ccarcinomasxe2x80x9d refers to lesions that are cancerous. Examples include malignant melanomas, breast cancer, and colon cancer.
As used herein, the term xe2x80x9cneoplasmxe2x80x9d refers to both precancerous and cancerous lesions.
Compounds useful in the methods of this invention may be formulated into compositions together with pharmaceutically acceptable carriers for oral administration in solid or liquid form, or for rectal administration, although carriers for oral administration are most preferred.
Pharmaceutically acceptable carriers for oral administration include capsules, tablets, pills, powders, troches and granules. In such solid dosage forms, the carrier can comprise at least one inert diluent such as sucrose, lactose or starch. Such carriers can also comprise, as is normal practice, additional substances other than diluents, e.g., lubricating agents such as magnesium stearate. In the case of capsules, tablets, troches and pills, the carriers may also comprise buffering agents. Carriers such as tablets, pills and granules can be prepared with enteric coatings on the surfaces of the tablets, pills or granules. Alternatively, the enterically coated compound can be pressed into a tablet, pill, or granule, and the tablet, pill or granules for administration to the patient. Preferred enteric coatings include those that dissolve or disintegrate at colonic pH such as shellac or Eudraget S.
Pharmaceutically acceptable carriers include liquid dosage forms for oral administration, e.g. pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents commonly used in the art, such as water. Besides such inert diluents, compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring and perfuming agents.
Pharmaceutically acceptable carriers for rectal administration are preferably suppositories that may contain, in addition to the compounds of Formula I, excipients such as cocoa butter or a suppository wax.
The pharmaceutically acceptable carrier and compounds of this invention are formulated into unit dosage forms for administration to a patient. The dosage levels of active ingredient (i.e. compounds of this invention) in the unit dosage may be varied so as to obtain an amount of active ingredient effective to achieve lesion-eliminating activity in accordance with the desired method of administration (i.e., oral or rectal). The selected dosage level therefore depends upon the nature of the active compound administered, the route of administration, the desired duration of treatment, and other factors. If desired, the unit dosage may be such that the daily requirement for active compound is in one dose, or divided among multiple doses for administration, e.g., two to four times per day.
The pharmaceutical compositions of this invention are preferably packaged in a container (e.g. a box or bottle, or both) with suitable printed material (e.g. a package insert) containing indications, directions for use, etc.
The dose of the compounds (I) for use in the present invention may vary in accordance with, for example, the administration routes, and the ages, weights and conditions of the patients. For example, when administered in an injection preparation, it is usually in the range of about 0.0001-0.5 mg/kg/day, preferably in the range of about 0.0005-0.1 mg/kg/day. When administered in an oral preparation, it is usually in the range of about 0.001-30 mg/kg/day, preferably in the range of about 0.05-10 mg/kg/day.
Among the compounds (1) useful in this present invention, a group of the formula xe2x80x94COOR3 is ones wherein R3 is a hydrogen atom, or an ester residue such as an aryl-lower alkyl group (e.g., benzyl, nitrobenzyl, a protected or unprotected amino-benzyl, a lower alkoxybenzyl), a lower alkyl group (e.g., methyl, ethyl, propyl, butyl), a cyclo-lower alkyl group (e.g., cyclopentyl), or a tri-lower alkylsilyl-lower alkyl group (e.g., trimethylsilylmethyl, tert-butyldimethylsilylmethyl). When R2 is a group xe2x80x94CON(R4)(R5), a group of the formula xe2x80x94N(4)(R5) is, for example, a substituted or unsubstituted nitrogen-containing 5 or 6-membered aliphatic heterocyclic group (e.g., a hydroxy-lower alkylsubstituted piperazinyl group, a morpholino group, a pyrrolidinyl group, a piperidinyl group), or a substituted or unsubstituted amino group (e.g., an imidazolyl-substituted lower alkylamino group, a mono- or di-lower alkylamino group, amino group).
Ring A and Ring B of the compounds (I) useful in the practice of this invention are a benzene ring which may optionally have 1 to 4 substituents being the same or different, and such substituents of said Ring A and Ring B are, for example, a protected or unprotected hydroxy group, a lower alkylenedioxy group, a halogen atom, a lower alkyl group, a mono- or di-lower alkylcarbamoyloxy group, or a group of the formula R6xe2x80x94(CO)nxe2x80x94Oxe2x80x94(R6 is a substituted or unsubstituted lower alkyl group, a substituted or unsubstituted lower alkenyl group, a substituted or unsubstituted cyclo-lower alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted arylsulfonyl group, or a substituted or unsubstituted heterocyclic group, and n is 0 or 1).
More particularly, Ring A of the compound (I) is a benzene ring which may optionally have 1 to 4 substituents being the same or different, and such substituents of Ring A are, for example, a protected or unprotected hydroxy group, a lower alkylenedioxy group, a halogen atom, a lower alkyl group, a mono- or di-lower alkylcarbamoyloxy group, or a group of the formula of the formula R6xe2x80x94(CO)nxe2x80x94Oxe2x80x94 (R6 is a substituted or unsubstituted lower alkyl group, a substituted or unsubstituted lower alkenyl group, a substituted or unsubstituted cyclo-lower alkyl group, a substituted or unsubstituted aryl group, a substituted or unsubstituted arylsulfonyl group, or a substituted or unsubstituted heterocyclic group, and n is 0 or 1). Ring B of the compound (I) of the present invention is a benzene ring which may optionally have 1 to 4 substituents being the same or different, and such substituents of Ring B are, for example, a protected or unprotected hydroxy group, a lower alkoxy group, a lower alkyl group, a halogen atom, or a lower alkylenedioxy group.
The suitable examples of Ring A and Ring B of the compounds of the present invention are those wherein Ring A is a benzene ring of the formula:
and Ring B is a benzene ring of the formula:
wherein A1 and A2 are the same or different and each a member selected from a hydrogen atom, a protected or unprotected hydroxy group, a lower alkylene-dioxy group, a halogen atom, a lower alkyl group, a mono- or di-lower alkyl-carbamoyloxy group, and a group of the formula R6xe2x80x94(CO)nxe2x80x94Oxe2x80x94(R6 and n are the same as defined above), B1, B2 and B3 are the same or different and each a member selected from a protected or unprotected hydroxy group, a lower alkoxy group, a lower alkyl group, a halogen atom and a lower alkylenedioxy group.
When Ring A and/or Ring B have a substituent of the formula R6xe2x80x94CO)nxe2x80x94Oxe2x80x94R6 is, for example, (1) a lower alkyl group which may optionally be substituted by 1 to 2 groups selected from a 5- to 10-membered heteromonocyclic or heterobicyclic group being optionally substituted by 1 to 4 groups selected from a hydroxy-substituted lower alkyl group, a lower alkyl group, an oxo group and a lower alkoxycarbonyl group; a 6- to 10-membered monocyclic or bicyclic aryl group being optionally substituted by 1 to 4 groups selected from a lower alkylenedioxy group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkoxy group, a sulfamoyl group, a carbamoyl group, a nitro group, a protected or unprotected amino group, a phenyl group, a halogen atom, a mono-lower alkylamino group, a di-lower alkylamino group, a lower alkyl-piperazinocarbonyl group, a hydroxy-substituted lower alkyl group and a lower alkyl group; a cyano group; a carboxyl group; a mono- or di-lower alkylamino group; a lower alkoxy-substituted lower alkoxy group; a lower alkoxy group; a hydroxy group; a carbamoyl group; a lower alkoxycarbonyl group; a cyclo-lower alkyl group; and a benzoyl group, or (2) a 5- to 10-membered heteromonocyclic or heterobicyclic group which may optionally be substituted by 1 to 4 groups selected from a lower alkyl group, a cyano group, a carboxyl group, a mono- or di-lower alkylamino group, a lower alkoxy-substituted lower alkyl group, a hydroxy group, a lower alkoxy group, a carbamoyl group, a lower alkoxycarbonyl group and a nitro group.
The 6- to 10-membered monocyclic or bicyclic aryl group is, for example, a phenyl group, a naphthyl group, etc., and the 5- to 10-membered heteromonocyclic or heterobicyclic group is, for example, a pyridyl group, a pyrimidinyl group, a pyrazinyl group, a piperidyl group, a piperazinyl group, a pyrrolidinyl group, an isoquinolyl group, a quinolyl group, a tetrazolyl group, a thienyl group, a furyl group, a morpholino group, a pyrrolyl group, a benzimidazolyl group, an imidazolyl group, a quinazolyl group, a phtalazinyl group, etc.
Among Ring A and Ring B, more preferable examples of Ring A are a benzene ring of the formula:
and more preferable examples of Ring B are a benzene ring of the formula:
wherein A1, A2, B1, B2 and B3 are the same as defined above.
Besides, among Ring A and Ring B, the most preferable examples of Ring
A are a benzene ring of the formula:
and the most preferable examples of Ring B are a benzene ring of the formula:
wherein A1, A2, B1, B2 and B3 are the same as defined above.
Suitable examples of the substituents (A1 and A2) of Ring A are, for example, a protected or unprotected hydroxy group; a lower alkoxy group which may optionally be substituted by a group selected from a lower aklene-dioxyphenyl group, a benzimidazolyl group, a lower alkyl-substituted imidazolyl group, a cyano group, a carboxyl group, a pyridyl group, an N-oxo-pyridyl group, a pyridyl group being substituted by a hydroxy-substituted lower alkyl group, a pyrrolidinyl group, an isoquinolyl group, a pyrimidinyl group, a pyrazinyl group, a quinazolyl group, a phthalazinyl group, a lower alkoxycarbonyl-substituted piperidyl group, a piperidyl group, a quinolyl group, a tetrazolyl group, a thienyl group, a furyl group, a pyrrolyl group being substituted by a lower alkyl group and a lower alkoxycarbonyl group, a mono-or di-lower alkyl amino group, a lower alkoxy-substituted lower alkoxy group, a lower alkoxy group, a hydroxy group, a carbamoyl group, a lower alkoxycarbonyl group, a cyclo-lower alkyl group, a hydroxy-lower alkyl group-substituted phenyl group, a carboxy-substituted phenyl group, a lower alkoxycarbonyl group-substituted phenyl group, a benzoyl group, a mono- or di-lower alkoxy-substituted phenyl group, a nitro-substituted phenyl group, a naphthyl group, a mono- or di-halogenophenyl group, a carbamoyl-substituted phenyl group, a sulfamoyl-substituted phenyl group, a phenyl group being substituted by one or two protected or unprotected amino groups, a biphenylyl group, a phenyl group being substituted by a halogen atom and a nitro group, a di-lower alkylamino-substituted phenyl group, and a lower alkyl-substituted phenyl group; a lower alkylenedioxy group; a halogen atom; a lower alkyl group; a cyclo-lower alkoxy group; a pyridyloxy group; a lower alkenyloxy group; a morpholinocarbonyloxy group; a lower alkyl-substituted piperazinylcarbonyloxy group; a pyrrolylcarbonyloxy group being substituted by a lower alkyl group and a nitro group; a pyrrolylcarbonyloxy group; a mono- or di-lower alkylcarbamoyloxy group; a lower alkyl-substituted phenylsulfonyloxy group; and a benzoyloxy group.
When R1 of the present compounds (I) is a substituted or unsubstituted aryl group, the aryl group is, for example, a 6- to 14-membered partially saturated or unsaturated monocyclic, bicyclic or tricyclic aryl group. The monocyclic aryl group is, for example, a phenyl group, a cyclohexadienyl group, a cyclohexenyl group, etc. The bicyclic aryl group is, for example, a naphthyl group, an indenyl group, an indanyl group, an azulenyl group, etc. The tricyclic aryl group is, for example, a fluorenyl group, a phenanthrenyl group, an anthracenyl group, etc.
When R1 of the present compounds (I) is a substituted or unsubstituted heterocyclic group, the heterocyclic group is, for example, a 5- to 12-membered partially saturated or unsaturated heteromonocyclic or heterobicyclic group, such as a 5- to 12-membered partially saturated or unsaturated aromatic heteromonocyclic or heterobicyclic group, or a 5- to 12-membered aliphatic heteromonocyclic or heterobicyclic group.
The 5- to 12-membered aromatic heteromonocyclic or heterobicyclic group is preferably a 5- to 10-membered aromatic heteromonocyclic or heterobicyclic group having 1 to 4 heteroatoms selected from a nitrogen atom, an oxygen atom and a sulfur atom, for example, a pyranyl group, an indazolyl group, a benzotriazolyl group, a pyrrolyl group, an imidazolyl group, a furyl group, a thienyl group, a thiazolyl group, an isoxazolyl group, an oxazolyl group, an oxazolinyl group, a pyrazolyl group, a phthalazinyl group, a quinazolinyl group, a thienopyrimidinyl group, a pyridyl group, a pyrimidinyl group, a pyridazinyl group, a pyrazinyl group, a triazinyl group, a tetrazolyl group, a quinolyl group, an isoquinolyl group, a quinoxalinyl group, an indolyl group, a benzothienyl group, a benzothiazolyl group, a benzoxazolyl group, or a benzimidazolyl group, and a partially saturated group of these groups.
The 5- to 12-membered aliphatic heteromonocyclic or heterobicyclic group is preferably a 5- to 10-membered aliphatic heteromonocyclic group having 1 to 4 heteroatoms selected from a nitrogen atom, an oxygen atom and a sulfur atom, for example, a piperazinyl group, a pyrrolidinyl group, a piperidyl group, a pyrazolidinyl group, a quinuclidinyl group, a thiomorpholino group, a morpholino group, a hexahydropyrimidinyl group, a tetrahydrofuranyl group, a tetrahydropyranyl group, and a dioxanyl group.
The substituent of the lower alkyl group for R1 of the present compounds (I) is, for example, a piperidyl group, a pyridyl group, an imidazolyl group, a lower alkyl-substituted piperidyl group, a furyl group, a morpholino group, a tetrahydrofuryl group, a dihydropyridyl group being substituted by a lower alkyl group and an oxo group, a piperazinyl group, a lower alkoxycarbonyl-substituted piperazinyl group, a cyclo-lower alkyl group, a phenyl group, a lower alkylenedioxyphenyl group, a lower alkoxycarbonyl group, a hydroxy group, a hydroxy-substituted lower alkoxy group, a caoxyl group, a lower alkoxy group, a protected or unprotected amino group, a carbamoyl group, a di-lower alkylamino group, and a pyridylcarbonyloxy group.
The lower alkyl group for R1 may optionally have 1 to 3 substituents being the same or different, which are selected from the above groups.
The substituent of the cyclo-lower alkyl group for R1 of the present compounds (I) is, for example, a lower alkoxycarbonyl group, a hydroxy group, a carboxyl group, a lower alkyl group, a lower alkoxy group, a hydroxy-substituted lower alkyl group, or a protected or unprotected amino group.
The cyclo-lower alkyl group for R1 may optionally have 1 to 3 substituents being the same or different, which are selected from the above groups.
The substituent of the aryl group for R1 of the present compounds (I) is, for example, a halogen atom, a mono- or di-lower alkylamino group, a morpholino group, a lower alkyl-substituted pyrimidinyl group, a lower alkyl-substituted pyrazolyl group, a hydroxy-substituted lower alkyl group, a protected or unprotected amino group, a lower alkanoyl-substituted amino group, a lower alkoxy group, a lower alkyl group, a protected or unprotected hydroxy group, a carboxy-substituted lower alkyl group, a lower alkoxy-carbonyl-substituted lower alkyl group, a lower alkoxycarbonyl-substituted lower alkoxy group, a carbamoyl group, a carboxyl group, a lower alkylthio group, a lower alkoxycarbonyl group, a nitro group, a trihalogeno-lower alkyl group, a morpholinodarbonyl group, a carboxy-substituted lower alkoxy group, a di-(lower alkylsulfonyl)amino group, a morpholino-lower alkylcarbamoyl-substituted lower alkoxy group, a sulfamoyl group, a lower alkyl group being substituted by a protected or unprotected amino group, an amino group being substituted by a lower alkyl group and a protecting group for amino group, a lower alkylenedioxy group, a carbamoyl group being substituted by a protected or unprotected amino group, a lower alkylsulfinyl group, and a lower alkyl-sulfonyl group.
The aryl group for R1 may optionally have 1 to 4 substituents being the same or different, which are selected from the above groups.
The substituent of the heterocyclic group for R1 of the present compounds (I) is, for example, a hydroxy group, a halogen atom, a lower alkyl group, a phenyl-substituted lower alkyl group, a hydroxy-substituted lower alkyl group, an oxo group, a lower alkoxy group, a protected or unprotected amino group, a mono- or di-lower alkylamino group, a phenyl-lower alkoxy-carbonyl group, a lower alkoxycarbonyl group, a carboxyl group, and a carbamoyl group.
The heterocyclic group for R1 may optionally have 1 to 4 substituents being the same or different, which are selected from the above groups.
The substituent of the amino group optionally having 1 or 2 substituents for R1 of the present compounds (I) is, for example, a protecting group for amino group, a pyridyl group, a lower alkanoyl group, a lower alkyl group, a hydroxy-substituted lower alkyl group, a phenyl group, a lower alkanoyloxy-substituted lower alkyl group, and a trihalogeno-lower alkanoyl group, which are the same or different.
When the present compounds (I) have a protected amino group, the protecting group for amino group is, for example, a substituted or unsubstituted lower alkoxycarbonyl group, a lower alkanoyl group, etc., such as a benzyloxycarbonyl group, a 4-methoxybenzyloxycarbonyl group, a 9-fluorenylmethyloxy-carbonyl group, a tert-butoxycarbonyl group, a 2,2,2-trichloroethyloxycarbonyl group, a formyl group, an acetyl group, a propionyl group, and a butyryl group. Among these groups, the preferable one is an aryl-substituted lower alkoxy-carbonyl group and an unsubstituted lower alkoxycarbonyl group, for example, a benzyloxycarbonyl and a tert-butoxycarbonyl group.
When the present compounds (I) have a protected hydroxy group, the protecting group for hydroxy group is a conventional protecting group such as a substituted or unsubstituted aryl-lower alkyl group, and an acyl group. Among these groups, the preferable one is, for example, an unsubstituted aryl-lower alkyl group (e.g., benzyl, phenethyl), and an acyl group (e.g., formyl, acetyl, propionyl, malonyl, acryloyl, benzoyl).
Among the desired compounds (I) of the present invention, the preferable compounds are compounds of the formula (I) wherein the aryl group is a phenyl group, an indanyl group or a naphthyl group, the heterocyclic group is a piperazinyl group, a pyranyl group, a morpholino group, an indazolyl group, a pyrrolidinyl group, an indolyl group, a benzotriazolyl group, a pyrazinyl group, a pyridyl group, a thiomorpholino group, a pyrrolyl group, a quinolyl group, an isoquinolyl group, a phthalazinyl group, an isoxazolyl group, or a piperidyl group, and the nitrogen-containing aliphatic heterocyclic group is a piperazinyl group or a morpholino group.
The more preferable compounds of the present invention are compounds of the formula (I) wherein Ring A is a benzene ring of the formula:
(A1 and A2 are the same or different and each a member selected from a protected hydroxy group; a lower alkoxy group; a pridyl-lower alkoxy group; a hydroxy-lower alkyl group-substituted pyridyl-lower alkoxy group; an N-oxopyridyl-lower alkoxy group; a pyrazinyl-lower alkoxy group; a quinolyl-lower alkoxy group; a lower alkoxy group being substituted by an amino-substituted phenyl group; a lower alkoxy group being substituted by a mono or di-lower alkylamino-substituted phenyl group; a lower alkoxy group being substituted by a lower alkoxy-substituted phenyl group; a lower alkoxy group being substituted by a hydroxy-lower alkyl group-substituted phenyl group; a lower alkoxy group being substituted by a carboxy-substituted phenyl group; and an isoquinolyl-lower alkoxy group), and Ring B is a benzene ring of the formula:
(B1, B2 and B3 are the same or different and each a member selected from a halogen atom, a lower alkyl group, and a lower alkoxy group), and R1 is a phenyl group optionally being substituted by a protected or unprotected amino group, or a pyridyl group optionally being substituted by a protected or unprotected amino group, or a morpholino group, and R2 is a lower alkoxycarbonyl group or a phenyl-lower alkoxycarbonyl group.
Among the desired compounds (I) of the present invention, more preferable compounds are compounds of the formula (I) wherein Ring A is a benzene ring of the formula:
and Ring B is a benzene ring of the formula:
(R6 is (1) a lower alkyl group which may optionally be substituted by a group selected from a pyrrolyl group optionally being substituted by a lower alkyl group or a lower alkoxycarbonyl group; a pyridyl group optionally being substituted by a hydroxy-lower alkyl group; a thienyl group; an N-oxopyridyl group; a pyrazinyl group; a phenyl group optionally being substituted by 1 to 3 groups being the same or different, and selected from a carboxyl group, a lower alkoxycarbonyl group, a nitro group, an amino group, a mono- or di-lower alkylamino group, a phenyl group, a halogen atom, a lower alkoxy group, a hydroxy-substituted lower alkyl group and a lower alkyl group, a naphthyl group; a quinolyl group; an isoquinolyl group; a benzimidazolyl group; and a cyclo-lower alkyl group, or (2) a pyrrolyl group optionally being substituted by a group selected from a lower alkyl group and a nitro group, A2 is a hydrogen atom or a lower alkoxy group, B1, B2 and B3 are the same or different and each a halogen atom, a lower alkyl group, or a lower alkoxy group, and n is 0 or 1), and R1 is a phenyl group, a phenyl group being substituted by a protected or unprotected amino group, or a morpholino group.
Other preferable compounds of the present invention are compounds of the formula (I) wherein Ring A is a benzene ring of the formula:
and Ring B is a benzene ring of the formula:
(A1 is a protected or unprotected hydroxy group, or a lower alkoxy group being substituted by a group selected from a pyridyl group, a hydroxy-lower alkyl group-substituted pyridyl group, an N-oxopyridyl group, a pyrazinyl group, an amino-substituted phenyl group, a mono- or di-lower alkylamino-substituted phenyl group, a lower alkoxy-substituted phenyl group, a hydroxy-lower alkyl group-substituted phenyl group, an isoquinolyl group and a quinolyl group, B1, B2 and B3 are the same or different and each a halogen atom, a lower alkyl group and a lower alkoxy group), and R1 is a phenyl group being substituted by a protected or unprotected amino group.
Among the desired compounds (I) of the present invention, pharmaceutically preferable compounds are compounds of the formula (I-A):
wherein Ring A and Ring B are the same or different and each a substituted or unsubstituted benzene ring, R1A is a substituted or unsubstituted aryl group or substituted or unsubstituted heterocyclic group, and R3 is a hydrogen atom or an ester residue, or a pharmaceutically acceptable salt thereof.
Examples of the above compounds are compounds of the formula (I-A) wherein Ring A and Ring B are the same or different and each a benzene ring having optionally 1 to 4 substituents selected from
(i) a hydroxy group;
(ii) a halogen atom;
(iii) a lower alkyl group;
(iv) a cyclo-lower alkoxy group;
(v) a lower alkylenedioxy group;
(vi) a lower alkoxy group;
(vii) a lower alkoxy group having 1 to 3 substituents selected from a hydroxy group, a benzoyl group, a lower alkoxycarbonyl group, a carboxyl group, a mono- or di-lower alkylamino group, a lower alkoxy-lower alkoxy group, a lower alkoxy group, a phenyl group, a naphthyl group and a phenyl group having 1 to 3 substituents selected from a nitro group, a halogen atom, a phenyl group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkyl group, a lower alkoxy group, an amino group, a mono- or di-lower alkylamino group and a hydroxy-lower alkyl group; and
(viii) a lower alkoxy group being substituted by a 5- to 10-membered heterocyclic group having 1 to 4 heteroatoms selected from a nitrogen atom, an oxygen atom and a sulfur atom, and optionally 1 to 3 substituents selected from a carboxyl group, a lower alkoxycarbonyl group, a lower alkyl group, a hydroxy-substituted lower alkyl group, a nitro group and an oxo group, R1A is a phenyl group; a phenyl group having 1 to 4 substituents selected from a protected or unprotected amino group, a halogen atom, a mono- or di-lower alkylamino group, a morpholino group, a lower alkyl-substituted pyrimidinyl group, a lower alkyl-substituted pyrazolyl group, a hydroxy-substituted lower alkyl group, a lower alkanoyl-substituted amino group, a lower alkoxy group, a lower alkyl group, a protected or unprotected hydroxy group, a carboxyl-substituted lower alkyl group, a lower alkoxycarbonyl-substituted lower alkyl group, a lower alkoxycarbonyl-substituted lower alkoxy group, a carbamoyl group, a carboxyl group, a lower alkylthio group, a lower alkoxycarbonyl group, a nitro group, a trihalogeno-lower alkyl group, a morpholinocarbonyl group, a carboxyl-substituted lower alkoxy group, a di-lower alkylsulfonyl-substituted amino group, a morpholino-lower alkylcarbamoyl-substituted lower alkoxy group, an amino group being substituted by a lower alkyl group and a protecting group for amino group, a lower alkylenedioxy group, a carbamoyl group being substituted by a protected or unprotected amino group, a lower alkylsulfinyl group and a lower alkylsulfonyl group; or a 5- to 10-membered heterocyclic group having 1 to 4 heteroatoms selected from a nitrogen atom, an oxygen atom and a sulfur atom, said heterocyclic group having 1 to 4 substituents selected from a hydroxy group, a halogen atom, a lower alkyl group, a phenyl-substituted lower alkyl group, a hydroxy-substituted lower alkyl group, an oxo group, a lower alkoxy group, a protected or unprotected amino group, a mono- or di-lower alkylamino group, a phenyl-substituted lower alkoxycarbonyl group, a lower alkoxycarbonyl group, a carboxyl group and a carbamoyl group, and
R3 is a hydrogen atom or a lower alkyl group.
Another embodiment of the compounds useful in the practice of the present invention is an isoquinolinone derivative of the formula (I-B):
wherein Ring A2 and Ring B2 are the same or different and each a benzene ring which may optionally be substituted by 1 to 4 groups selected from the group consisting of a protected or unprotected hydroxyl group; a lower alkylenedioxy group; a halogen atom; a lower alkyl group; a mono- or di-lower alkylcarbamoyloxy group; and a group of the formula:
R6Bxe2x80x94(CO)nxe2x80x94O
in which R6B is
(i) a lower alkyl group which may optionally have 1 or 2 substituents selected from the group consisting of a 5- to 12-membered heteromonocyclic or heterobicyclic group having optionally 1 to 4 substituents selected from the group consisting of a hydroxy-substituted lower alkyl group, a lower alkyl group, an oxo group and a lower alkoxycarbonyl group; a phenyl or naphthyl group having optionally 1 to 4 substituents selected from the group consisting of a protected or unprotected amino group, a lower alkylenedioxy group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkoxy group, a sulfamoyl group, a carbamoyl group, a nitro group, a phenyl group, a halogen atom, a mono-lower alkylamino group, a di-lower alkylamino group, a lower alkylpiperazinocarbonyl group, a hydroxy-substituted lower alkyl group and a lower alkyl group; a cyano group; a carboxyl group; a mono or di-lower alkylamino group; a lower alkoxy-substituted lower alkoxy group; a lower alkoxy group; a hydroxy group; a carbamoyl group; a lower alkoxycarbonyl group; a cyclo-lower alkyl group; and a benzoyl group,
(ii) a 5- to 12-membered heteromonocyclic or heterobicyclic group having optionally 1 to 4 substituents selected from the group consisting of a lower alkyl group, a cyano group, a carboxyl group, a mono- or di-lower alkylamino group, a lower alkoxy-substituted lower alkyl group, a hydroxy group, a lower alkoxy group, a carbamoyl group, a lower alkoxycarbonyl group and a nitro group,
(iii) a cyclo-lower alkyl group,
(iv) a lower alkenyl group, or
(v) a lower alkyl-substituted or unsubstituted phenylsulfonyl group, n is an integer of 0 or 1,
R1B is
(i) a hydrogen atom,
(ii) a lower alkyl group having optionally 1 to 3 substituents selected from the group consisting of a piperidyl group, a pyridyl group, an imidazolyl group, a lower alkyl-substituted piperidyl group, a furyl group, a morpholino group, a tetrahydrofuryl group, a dihydropyridyl group being substituted by a lower alkyl group and an oxo group, a piperazinyl group, a lower alkoxycarbonyl substituted-piperazinyl group, a cyclo-lower alkyl group, a phenyl group, a lower alkylenedioxy-phenyl group, a lower alkoxycarbonyl group, a hydroxyl group, a hydroxy-substituted lower alkoxy group, a carboxyl group, a lower alkoxy group, a protected or unprotected amino group, a carbamoyl group, a di-lower alkylamino group and a pyridylcarbonyloxy group,
(iii) a cyclo-lower alkyl group having optionally 1 to 3 substituents selected from the group consisting of a lower alkoxycarbonyl group, a hydroxy group, a carboxyl group, a lower alkyl group, a lower alkoxy group, a hydroxy-substituted lower alkoxy group and a protected or unprotected amino group,
(iv) an unsaturated or partially saturated 6- to 14-membered monocyclic, bicyclic or tricyclic aryl group having optionally 1 to 4 substituents selected from the group consisting of a halogen atom, a mono- or di-lower alkylamino group, a morpholino group, a lower alkyl-substituted pyrimidinyl group, a lower alkyl-substituted pyrazolyl group, a hydroxy-substituted lower alkyl group, a protected or unprotected amino group, a lower alkanoyl-substituted amino group, a lower alkoxy group, a lower alkyl group, a protected or unprotected hydroxy group, a carboxy-substituted lower alkyl group, a lower alkoxycarbonyl-substituted lower alkyl group, a lower alkoxycarbonyl-substituted lower alkoxy group, a carbamoyl group, a carboxyl group, a lower alkylthio group, a lower alkoxycarbonyl group, a nitro group, a trihalogeno-lower alkyl group, a morpholinocarbonyl group, a carboxy-substituted lower alkoxy group, a di-lower alkylsulfonylamino group, a morpholino-lower alkyl carbamoylsubstituted lower alkyl group, a sulfamoyl group, a carbamoyl group being optionally substituted by a protected or unprotected amino group, a lower alkylsulfinyl group and a lower alkylsulfonyl group,
(v) a 5- to 12-membered aromatic or aliphatic heteromonocyclic or heterobicyclic group having 1 to 4 substituents selected from the group consisting of a hydroxy group, a halogen atom, a phenyl-substituted lower alkyl group, a hydroxy-substituted lower alkyl group, an oxo group, a lower alkoxy group, a protected or unprotected amino group, a mono- or di-lower alkylamino group, a phenyl-lower alkoxycarbonyl group, a lower alkoxycarbonyl group, a carboxyl group and a carbamoyl group, or
(vi) an amino group having optionally 1 or 2 substituents selected from the group consisting of a protecting group for amino group, a pyridyl group, a lower alkanoyl group, a lower alkoxy group, a hydroxy-substituted lower alkyl group, a phenyl group, a lower alkanoyloxy-substituted lower alkyl group and a trihalogeno-lower alkanoyl group, R2B is a group of the formula: xe2x80x94COOR3B or a group of the formula: xe2x80x94CON(R4B)(R5B) R3B is a hydrogen atom, a lower alkyl group, a tri-lower alkylsilyl group or a phenyl-lower alkyl group, and a group of the formula: xe2x80x94N(R4B)(R5B) is a hydroxy-lower alkyl-substituted piperazinyl group, a morpholino group, a pyrrolidinyl group, an imidazolyl-substituted lower alkylamino group or a mono, or di-lower alkylamino group, provided that when R1B is one of the groups of the above-mentioned (i) or (ii), then at least one of Ring A2 and Ring B2 is a benzene ring which is substituted by two or more lower alkoxy groups.
Examples of the pharmaceutically preferable compounds are as follows.
6-methoxy-3-methoxycarbonyl-2-morpholino-7-(4-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)isoquinolinone;
6-methoxy-3-methoxycarbonyl-2-morpholino-7-(3-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
6-methoxy-3-methoxycarbonyl-2-morpholino-7-(2-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
6-methoxy-3-methoxycarbonyl-2-phenyl-4-(3,4,5-trimethoxyphenyl)-7-(4-pyridylmethyloxy)-1(2H)-isoquinolinone;
6-methoxy-3-methoxycarbonyl-2-phenyl-4-(3,4,5-trimethoxyphenyl)-7-(3-pyridylmethyloxy)-1(2H)-isoquinolinone; or
6-methoxy-3-methoxycarbonyl-2-phenyl-4-(3,4,5-trimethoxyphenyl)-7-(2-pyridylmethyloxy)-1(2H)-isoquinolinone.
Among the desired compounds (I) of the present invention, other pharmaceutically preferable compounds are as follows.
2-(4-aminophenyl)-6-methoxy-3-methoxycarbonyl-7-(2-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3-aminobenzyloxy)-6-methoxy-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-6-methoxy-3-methoxycarbonyl-7-(4-pyridylmethyl-oxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(2-benzimidazolylmethyloxy)-6-methoxy-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3,5-diaminobenzyloxy)-6-methoxy-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4(4-bromo-3,5-dimethoxyphenyl)-6-methoxy-3-methoxycarbonyl-7-(2-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-bromo-3,5-dimethoxyphenyl)-6-methoxy-3-methoxycarbonyl-7-(3-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4(4-bromo-3,5-dimethoxyphenyl)-6-methoxy-3-methoxycarbonyl-7-(4-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-(2-pyridylmethyloxy)-4-(3,4,5-methoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-(3-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-(4-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(2,5-methoxybenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3,5-dimethoxybenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-bromo-3,5-dimethoxyphenyl)-3-methoxycarbonyl-7-(2-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-bromo-3,5-dimethoxyphenyl)-3-methoxycarbonyl-7-(3-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-bromo-3,5-dimethoxyphenyl)-3-methoxycarbonyl-7-(4-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3-aminobenzyloxy)-4-(4-bromo-3,5-dimethoxy-phenyl)-3-methoxycarbonyl-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-chloro-3,5-dimethoxyphenyl)-3-methoxy-carbonyl-7-(2-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-chloro-3,5-dimethoxyphenyl)-3-methoxy-carbonyl-7-(3-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3-dimethylaminobenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-pyrazinylmethyloxy-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-chloro-3,5-dimethoxyphenyl)-3-methoxy-carbonyl-7-(4-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3,5-diaminobenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2(4-aminophenyl)-7-(6-hydroxymethyl-2-pyridyimethyloxy)-3-methoxy-carbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(4-carboxybenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3-carboxybenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-[4-(4-methylpiperazinylcarbonyl)benzyloxy]-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-[3-(4-methylpiperazinylcarbonyl)benzyloxy]-4-(3,4,5-trimethoxyphenyl)-1(2H-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-[3-(methylamino)benzyloxy]-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(2-hydroxymethylbenzyloxy)-3-methoxycarbonyl-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-7-(N-oxo-2-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycabonyl-8-(2-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-8-(3-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-8-(4-pyridylmethyloxy)-4-(3,4,5-trimethoxyphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-chloro-3,5-dimethoxyphenyl)-7-(6-hydroxymethyl-2-pyridylmethyloxy)-3-methoxycarbonyl-1(2H)-isoquinolinone;
2-(4-aminophenyl)-4-(4-chloro-3,5-dimethoxyphenyl)-3-methoxycarbonyl-7-pyrazinylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-7-(4-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-7-(3-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methyl-phenyl)-7-(2-pyridylmethyloxy)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3,5-diaminobenzyloxy)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(6-hydroxymethyl-2-pyridylmethyloxy)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(3-methylaminobenzyloxy)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-1(2H)-isoquinolinone;
2-(4-aminophenyl)-7-(2-hydroxymethylaminobenzyloxy)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-1(2H)-isoquinolinone; or
2-(4-aminophenyl)-3-methoxycarbonyl-4-(3,5-dimethoxy-4-methylphenyl)-7-(2-pyrazinylmethyloxy)-1(2H)-isoquinolinone.
When the desired compound (I) of the present invention has an asymmetric carbon atom at the substituents of Ring A and Ring B and/or at R1, it may exist in the form of an optically active isomer thereof owing to said asymmetric carbon atom thereof, and the present invention also includes these optical isomers and a mixture thereof.
The present compounds (I) can clinically be used either in the free form or in the form of a pharmaceutically acceptable salt thereof. The pharmaceutically acceptable salt includes a salt with an inorganic acid such as hydrochloride, sulfate or hydrobromide, or a salt with an organic acid such as acetate, fumarate, oxalate, citrate, methanesulfonate, tosylate, or maleate. The compounds (I) having a substituent such as a carboxyl group may clinically be used in the form of a salt with a base such as an alkali metal salt (e.g., sodium salt, potassium salt) or an alkaline earth metal salt (e.g., calcium salt) as well.
The desired compound (I) or a salt thereof includes either intramolecular salt or an additive thereof, and solvates or hydrates thereof.
Compounds of Formula I may be prepared by any suitable method known in the art or by the following processes that are set forth in PCT/JP98/00715. In the methods below, R1, R2 etc. are as defined in Formula I above, unless otherwise indicated.
The desired compounds (I) useful in the present invention may be prepared by the following Processes A, B, and C.
Process A
The desired compounds (I) useful in the present invention can be prepared by reacting an isocoumarin derivative of the formula (II):
wherein the symbols are the same as defined above, or a salt thereof, with an amine compound of the formula (III):
R1xe2x80x94NH2
wherein the symbol is the same as defined above, or a salt thereof.
Process B
The desired compounds (I) useful in the present invention can be prepared by subjecting an isocoumarin derivative of the formula (II)
wherein the symbols are the same as defined above, or a salt thereof, to hydrolysis to give a compound of the formula (IV):
wherein the symbols are the same as defined above, and reacting the compound (IV) with an amine compound of the formula (III):
R1xe2x80x94NH2
wherein the symbol is the same as defined above, or a salt thereof. The compound of the formula (IV) may exist in a solution in equilibration as follows.
Process C
The desired compounds (I) useful in the present invention can be prepared subjecting a benzoylbenzamide, compound of the formula (V):
wherein the symbols are the same as defined above, or a salt thereof, to intramolecular cyclization reaction, to give a compound of the formula (VI):
wherein the symbols are the same as defined above, and subjecting the compound (VI) to dehydration reaction.
The compound (I) obtained by Process A, B or C may, if necessary, be converted into a pharmaceutically acceptable salt thereof.
The above Processes A, B and C can be carried out as follows.
Process A
The reaction between the isocoumarin derivative (II) and the amine compound (III) or a salt thereof is carried out in a solvent or without a solvent. The solvent includes, for example, 1,3-dimethyl-2-imidazolidinone (DMI), dimethylformamide, dimethylsulfoxide, ethylene glycol, N-methylpyrrolidone, xylene, dichloroethane, etc. The reaction is carried out at 20-150xc2x0 C., preferably at 40-130xc2x0 C.
Process B
The hydrolysis of the isocoumarin derivative (II) is carried out in the presence of a strong base in a solvent. The strong base includes, for example, an alkali metal hydroxide (e.g., sodium hydroxide, potassium hydroxide, lithium hydroxide, etc.), an alkali metal carbonate (e.g., sodium carbonate, potassium carbonate, etc.), etc. The solvent includes, for example, water, or a mixture of water and methanol, ethanol, tetrahydrofuran, dioxane, dimethylformamide, etc. The reaction is carried out at 0-80xc2x0 C., preferably at 5-60xc2x0 C.
The reaction between the compound (IV) and the amine compound (III) is carried out in the presence or absence of an acid acceptor in a suitable solvent or without a solvent. The acid acceptor includes N-methylmorpholine, triethylamine, pyridine, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, etc. The solvent may be any solvents used in the above Process A which does not disturb the reaction. The reaction is carried out at 20-140xc2x0 C., preferably at 30-100xc2x0 C.
Process C
The intramolecular cyclization reaction of the benzoylbenzamide compound (V) is carried out in the presence or absence of a base in a solvent. The base includes, for example, an organic base (e.g., 1,5-diazabicyclo[4.3.0]-non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), etc.), or an organic base (e.g., sodium methoxide, potassium tert-butoxide, sodium hydride, n-butyl lithium, lithium diisopropyl amide, etc.), and these bases are usually used in an amount of 0.5-5 equivalents, preferably in an amount of 1-2 equivalents, to 1 equivalent of the compound (V). The solvent includes tetrahydrofuran, dimethylformamide, dioxane, dimethoxyethane, benzene, toluene, pyridine, etc., but may be any solvent used in the above Process A which does not disturb the reaction. The reaction is carried out at xe2x88x9250-100xc2x0 C., preferably at xe2x88x9220-80xc2x0 C.
The dehydration reaction of the compound (VI) is carried out in the presence of an acidic catalyst in a solvent. The acidic catalyst includes a sulfonic acid compound (e.g., p-toluenesulfonic acid, methanesulfonic acid, etc.), a carboxylic acid compound (e.g., acetic acid, trifluoroacetic acid, etc.), an inorganic acid compound (e.g., hydrogen chloride, hydrogen bromide, sulfuric acid, etc.), and a Lewis acid (e.g., boron trifluoride ethyl ether, aluminum chloride, etc.), and these acidic catalysts are usually used in an amount of 0.1-5 equivalent, preferably in an amount of 0.2-2 equivalents, to the amount of the compound (VI). The solvent includes, for example, chloroform, dioxane, benzene, toluene, methylene chloride, etc., but may be any solvent used in the above Process A which does not disturb the reaction. The reaction is carried out at 0-150xc2x0 C., preferably at 20-110xc2x0 C.
When R1 of the amine compound (III) used in the above Processes A and B is an amino group, or a group containing an amino group, these Processes A and B are preferably carried out after introducing a protecting group such as a substituted or unsubstituted lower alkoxycarbonyl group (e.g., tert-butoxycarbonyl group, benzyloxycarbonyl group, etc.), and a lower alkanoyl group (e.g., formyl group, acetyl group, propionyl group, etc.) into said amino group.
The compound of formula (I) obtained in the above Processes A, B and C wherein the group xe2x80x94COOR3 is a carboxyl group, i.e., a compound of the formula (I-a):
wherein the symbols are the same as defined above, is converted into a compound of the formula (I-b):
wherein R31 is an ester residue and the other symbols are the same as defined above, by esterification reaction in a conventional manner. For example, the compound (I-b) is prepared by reacting the compound (I-a) with an esterifying agent in the presence or absence of an acid acceptor in a solvent. The acid acceptor includes, for example, an inorganic base (e.g., an alkali metal hydroxide, an alkali metal carbonate, etc.), and an organic base (e.g., N-methylmorpholine, triethylamine, pyridine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diaza-bicyclo[5.4.0]undec-7-ene (DBU), etc.). The esterifying agent includes, for example, a diazoalkane (e.g., diazomethane, diazoethane, etc.), a dialkyl sulfate (e.g., dimethyl sulfate, diethyl sulfate, etc.), an alkyl halide (e.g., methyl iodide, methyl bromide, ethyl bromide, etc.), a tri-lower alkylsilyldiazoalkane (e.g., trimethylsilyldiazomethane, etc.), an aryl-lower alkyl halide (e.g., benzyl chloride, benzyl bromide, etc.), etc. When a dialkyl sulfate, an alkyl halide or an aryl-lower alkyl halide is used as an esterifying agent, the acid acceptor is usually used in an amount of 1-5 equivalents, preferably in an amount of 1-2 equivalents, to 1 equivalent of the compound (1-a). The reaction is carried out at 0-60xc2x0 C., preferably at 5-40xc2x0 C. When a diazoalkane is used as an esterifying agent, the acid acceptor is usually used in an amount of 1-5 equivalents, preferably in an amount of 1-2 equivalents, to 1 equivalent of the compound (I-a). The reaction is carried out at 0-50xc2x0 C., preferably at 5-30xc2x0 C. The compound of the formula (1-a) wherein the group xe2x80x94COOR3 is a methoxycarbonyl group is prepared under moderate conditions by using trimethylsilyldiazomethane as an esterifying agent in the above reaction. The solvent includes, for example, water, an alcohol (e.g., methanol, ethanol, isopropyl alcohol, etc.), an ether (e.g., diethyl ether, tetrahydrofuran, dioxane, etc.), a ketone (e.g., acetone, methyl ethyl ketone, etc.), as ester (e.g., ethyl acetate, etc.), an aromatic hydrocarbon (e.g., benzene, toluene, etc.), a halogenated hydrocarbon (e.g., methylene chloride, chloroform, etc.), an amide (e.g., N,N-dimethylformamide, N,N-dimethylacetamide, etc.), a sulfoxide (e.g., dimethylsulfoxide, etch), or a mixture of these solvents.
In addition, the compound (I-b) is prepared by reacting the compound (I-a) with a lower alcohol (e.g., methanol, ethanol, propanol, butanol, etc.), or an aryl-lower alcohol (e.g., benzyl alcohol, phenethyl alcohol, etc.), under acidic conditions. The acid includes, for example, sulfuric acid, hydrogen chloride, p-toluenesulfonic acid, etc., which is usually used in an amount of 0.01-20 equivalents, preferably in an amount of 0.1-10 equivalents, to 1 equivalent of the compound (1-a). The reaction is preferably carried out in said alcohol with heating under reflux.
When the above-mentioned compound (I-a) has one or more carboxyl group or a mono-substituted or unsubstituted amino group except the 3-carboxyl group (xe2x95x90R2), said compound (I-a) can be converted into a corresponding compound of the formula (I-a) wherein said carboxyl group is esterified, or said amino group is converted into a mono- or di-lower alkylamino group, by reacting with the above-mentioned esterifying agent.
Besides, the compound (I) wherein the substituent R2 is a group of the formula xe2x80x94CON(R4)(R5), i.e., a compound of the formula (1-c):
wherein a group of the formula xe2x80x94N(R4)(R5) is a substituted or unsubstituted nitrogen-containing aliphatic heterocyclic group, or a substituted or unsubstituted amino group, and the other symbols are the same as defined above, is prepared by reacting the compound of the formula (I-a) with an amine compound of the formula (VII):
wherein the symbols are the same as defined above, in the presence of an condensing agent, or reacting a reactive derivative (e.g., an acid halide, an active amide, an active ester, a mixed acid anhydride, etc.) of the compound (I-a) with the amide compound (VII) in the presence or absence of a base in a solvent. The base includes an organic base (e.g., pyridine, 4-dimethylaminopyridine, N-methylmorpholine, triethylamine, N,N-dimethylaniline, N,N-diethylaniline, 1,8-diazabicyclo[5.4.0]undec-7-ene, etc., and an inorganic base (e.g., sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, etc.). The condensing agent includes, for example, 1,3-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (WSCI), propanephosphonic anhydride (PPA), etc. The solvent includes, for example, dimethylformamide, methylene chloride, tetrahydrofuran, dioxane, ethyl acetate, and 1,3-dimethyl-2-imidazolidinone, but may be any solvent used in the above Process A that does not disturb the reaction. The reaction is carried out at xe2x88x9220-60xc2x0 C., preferably at 5-40xc2x0 C.
The active ester of the compound (I-a) is preferably an ester of the compound (I-a) with N-hydroxysuccinmide, N-hydroxyphthalimide, 1-hydroxybenzotriazole, or p-nitrophenol.
The acid halide of the compound (I-a) is preferably an acid chloride, an acid bromide, etc.
The active amide of the compound (I-a) is preferably an amide of the compound (1-a) with imidazole, etc.
In the preparation of the desired compound (a), the starting compounds (I-a), (I-d), (I-f), (1-h), (II), (III), (IV), (V), (VI) and (VII) in the above Processes A to C as well as to Steps (a) to (d) disclosed hereinafter can be used as well as in the form of a salt thereof. The salt may be, for example, a salt with an alkali metal (e.g., sodium, potassium, lithium, etc.), a salt with an organic base (e.g., pyridine, triethylamine, N-methylmorpholine, etc.), a salt with an inorganic acid (e.g., hydrogen chloride, hydrogen bromide, sulfuric acid, etc.), or a salt with an organic acid (e.g., acetic acid, formic acid, oxalic acid, citric acid, malonic acid, etc.).
The desired compound (I) of the present invention can also be prepared by converting the substituents of Ring A and/or Ring B, or the substituents R1 and/or R2 into other substituents. The method for conversion reaction of these substituents; is selected in accordance with the kinds of the substituents to be required, and may be the following steps (a) to (t).
Step (a)
The compound of the formula (I-e):
wherein the symbols are the same as defined above, is prepared by reacting a compound of the formula (I-d):
wherein the symbols are the same as defined above, or a salt thereof, with a compound of the formula (VIII-a):
R6xe2x80x94COOH
wherein R6 is the same as defined above, or a reactive derivative thereof, or with a compound of the formula (VIII-b):
R6xe2x80x94X
wherein X is a leaving group, and R6 is the same as defined above.
The leaving group (X) of the compound (VIII-b) is, for example, a hydroxy group, a tifluoromethanesulfonyloxy group, a p-tosyloxy group, a methanesulfonyloxy group, or a halogen atom such as chlorine, bromine, iodine, etc.
The reaction between the compound (I-d) and the compound (VIII-a) is carried out in the presence of a condensing agent (e.g., 1,3-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, diethylphosphonic cyanide, diphenylphosphonic azide, etc.). The reaction between a reactive derivative of the compound (VIII-a) (e.g., an active ester such as N hydroxysuccinimide ester, N-hydroxyphthalimide ester, 1-hydroxybenzotriazole ester, or an acid halide such as acid chloride, acid bromide, etc.) and the compound (1-d) is carried out in the presence of an acid acceptor such as an alkali metal hydroxide (e.g., sodium hydrochloride), an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate), an alkali metal carbonate (e.g., sodium carbonate), or an organic base (e.g., triethylamine, pyridine, etc.), and if necessary, 4-dimethylaminopyridine (DMAP), etc. may be added to the reaction mixture in a catalytic amount. The reaction is carried out at 0-80xc2x0 C., preferably at 5-60xc2x0 C.
The reaction between the compound (I-d) and the compound (VIII-b) is carried out, for example, according to the method disclosed in Mitsunobu, et al. (cf., Synthesis, pp. 1-28, 1981), when the leaving group X is a hydroxy group. That is, the compound (I-d) and the compound (VIII-b) are reacted in the presence of diethyl azodicarboxylate (DEAD) and triphenylphosphine in a solvent such as tetrahydrofuran, dioxane, ethyl acetate, dimethylformamide, chloroform, methylene chloride, benzene, toluene, dimethoxyethane, etc. The reaction is carried out at 0-60xc2x0 C., preferably at 5-40xc2x0 C.
When the leaving group X of the compound (VIII-b) is a trifluoromethanesulfonyloxy group, p-tosyloxy group, methanesulfonyloxy group, or a halogen atom such as chlorine, bromine, iodine, etc., the reaction between the compound (I-d) and the compound (VIII-b) is carried out in the presence of a base. The reaction may also be carried out in the presence or absence of a base and/or a copper catalyst, The base includes, for example, an inorganic base such as an alkali metal hydride (e.g., sodium hydride), an alkali metal amide (e.g., sodium amide), an alkali metal alkoxide (e.g., sodium methoxide, potassium tert-butoxide), an alkali metal hydroxide (e.g., sodium hydroxide), an alkali metal carbonate (e.g., sodium carbonate), and an organic base such as N-methylmorpholine, triethylamine, pyridine, etc. The base is usually used in an amount of 1 to 5 equivalents, preferably in an amount of 1 to 2 equivalents, to 1 equivalent of the compound (I-d). When the substituent R1 is an amino group optionally having one substituent, or a substituent containing an amino group optionally having one substituent, it is preferable to carry out the reaction after introducing a protecting group such as a lower alkoxycarbonyl group (e.g., tert-butoxycarbonyl group), an aryl-lower alkoxycaTbonyl group (e.g., benzyloxycarbonyl group), or a lower alkanoyl group (e.g., formyl group, acetyl group, propionyl group) into said amino group.
The copper catalyst may be copper(I) iodide, copper(II) bromide, copper(0) powder, copper(1) oxide, copper(II) bromide, etc. The reaction is carried out at 10-160xc2x0 C., preferably at 20-120xc2x0 C.
The desired compound (I) wherein Ring A is a benzene ring being substituted by a group selected from a lower alkyl-substituted piperazinylcarbonyloxy group, and a mono. or di-lower alkylcarbamoyloxy group is prepared by reacting the compound (I-d) with phosgene or triphosgene, followed by reacting the resulting corresponding product (chloroformate compound) with a lower alkyl-substituted piperazine or a mono- or di-lower alkylamine in the presence or absence of a base (e.g., triethylamine, N-methylmorpholine, pyridine, 4-dimethylaminopyridine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), etc.). The reaction is carried out at 0-80xc2x0 C., preferably at 10-40xc2x0 C.
Step (b)
The compound of the formula (I-g):
wherein R12 is an amino-substituted lower alkyl group, an amino-substituted cyclo-lower alkyl group, an amino-substituted aryl group, an amino-substituted heterocyclic group, or an amino group, and the other symbols are the same as defined above, is prepared by removing a protecting group for amino group from a compound of the formula (I-f):
wherein R11 is a lower alkyl group being substituted by a protected amino group, a cyclo-lower alkyl group being substituted by a protected amino group, an aryl group being substituted by a protected amino group, a heterocyclic group being substituted by a protected amino group, or a protected amino group, and the other symbols are the same as defined above, or a salt thereof.
The removal of the protecting group is carried out by a conventional method such as acid-treatment, base-treatment, catalytic reduction, etc., which is selected according to the kinds of the protecting group to be removed. The reaction is carried out at 0-150xc2x0 C., preferably at 5-110xc2x0 C.
Step (c)
The compound of the formula (I-i):
wherein R13 is a substituted or unsubstituted lower alkyl group, and the other symbols are the same as defined above, is prepared by reacting a compound of 10 the formula (1-h):
wherein the symbols are the same as defined above, or a salt thereof, with a compound of the formula (IX):
R13xe2x80x94X1
wherein X1 is a halogen atom, and R13 is the same as defined above.
The reaction between the compound (I-h) and the compound (IX) is carried out in the presence of an acid acceptor. The acid acceptor is, for example, an alkali metal hydroxide (e.g., sodium hydroxide), an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate), an alkali metal carbonate (e.g., sodium carbonate), an alkali metal hydride (e.g., sodium hydride), or an organic base (e.g., triethylamine, pyridine, 1,8-diazabicyclo[5.4.0]-undec-7-ene, etc.). The reaction is carried out at 0-100xc2x0 C., preferably at 20-80xc2x0 C.,
Step (d)
The desired compound (I) wherein the substituent of Ring A and/or the substituent R1 are a substituent containing an esterified carboxyl group (e.g., a lower alkoxycarbonyl-substituted aryl group, a lower alkoxycarbonyl-substituted-lower alkyl group, a lower alkoxycarbonyl-substituted cyclo-lower alkyl group, a lower alkoxycarbonyl-substituted lower alkyl-substituted aryl group, a lower alkoxycarbonyl-substituted lower alkoxy-substituted aryl group, etc.) is prepared by subjecting a corresponding compound of the formula (I) wherein the substituent of Ring A and/or the substituent R1 are a substituent containing a free carboxyl group, to esterification reaction. The reaction is carried out in the same manner as in the esterification reaction of the compound (1-a) as mentioned above.
The Step (e)
The desired compound (I) wherein the substituent of Ring A and/or the substituent R1 are a substituent containing a free carboxyl group (e.g., a carboxy-substituted aryl group, a carboxy-substituted cyclo-lower alkyl group, a carboxy-lower alkyl-substituted aryl group, a carboxy-lower alkyl group, a carboxy-lower alkoxy-substituted aryl group, a carboxy-substituted lower alkoxy group, a carboxy-aryl-substituted lower alkyl group, etc.) is prepared by subjecting a corresponding compound of the formula (I) wherein the substituent of Ring A and/or the substituent R1 are a substituent containing an esterified carboxyl group, to desterification reaction, for example, hydrolysis with a base (e.g., sodium hydroxide), an acid-treatment with a trifluoroacetic acid, hydrogen chloride, hydrogen bromide, etc., or reduction under hydrogen atmosphere with palladium-black, or palladium-carbon, which is selected according to the kinds of the ester residue to be removed. Among these de-esterification reactions, the hydrolysis with a base is carried out at 5-70xc2x0 C., the acid-treatment is carried out at 5-80xc2x0 C., and the reduction is carried out at 10-40xc2x0 C.
Step (f)
The desired compound (I) wherein R1 is an aryl group being substituted by a protected or unprotected amino-substituted carbamoyl group, or an aryl group being substituted by a morpholinocarbonyl group is prepared by reacting a corresponding compound of the formula (I) wherein R1 is carboxy-substituted aryl group with an amine compound of the formula:
HN(Ra)(Rb)
wherein one of Ra and Rb is a hydrogen atom, and the other is a protected or unprotected amino group, or both combine at their termini together with the adjacent nitrogen atom to form a morpholino group, in the presence of a condensing agent. The condensing agent includes a conventional one which is usually used in the amido-bond formation reaction between a carboxylic acid and an amine, for example, 1,1-carbonyldiimidazole (CDI), DCC, WSCI, isobutyl chloroformate and N-methylmorpholine, etc. The reaction is carried out at 0-50xc2x0 C. When Ra or Rb of the product is a protected amino group, if necessary, said protecting group may be removed by a conventional method.
The desired compound (I) wherein the group R1 is an aryl group being substituted by a morpholino-lower alkylcarbamoyl-substituted lower alkoxy group is prepared by reacting a corresponding compound of the formula (I) wherein R1 is an aryl group being substituted by a carboxyl-substituted lower alkoxy group with a morpholino-lower alkylamine in the same manner as above.
The desired compound (I) wherein Ring A is a lower alkoxy-substituted benzene ring being substituted by a carbamoyl group is prepared by reacting a corresponding compound (I) wherein Ring A is a benzene ring being substituted by a carboxyl-substituted lower alkoxy group with ammonia in the same manner as above.
Step (g)
The desired compound (I) wherein the substituent of Ring A and/or the group R1 is a substituent containing an amino group (e.g., an amino-lower alkyl-substituted aryl group, an amino-substituted aryl-substituted lower alkyl group, etc.) is prepared by removing a lower alkanoyl group or a protecting group for amino group from a corresponding compound of the formula (I) wherein the substituent of Ring A and/or the group R1 is a substituent containing a mono-or di-lower alkanoylamino group or a protected amino group. The removal of said protecting group for amino group or said lower alkanoyl group is carried out by a conventional method (e.g., acid-treatment, base-treatment, catalytic reduction, etc.). The acid-treatment is carried out at 5-120xc2x0 C., the base-treatment is carried out at 5-40xc2x0 C., and the catalytic reduction is carried out at 10-40xc2x0 C.
Step (h)
The desired compound (I) wherein the substituent of Ring A and/or the group R1 is a substituent containing a heterocyclic group (e.g., piperazinyl group, piperidinyl group, or pyrrolidinyl group) is prepared by removing the N-substituent (i.e., a lower alkoxycarbonyl group, or an aryl-lower alkoxycarbonyl group) from a corresponding compound of the formula (I) wherein the substituent of Ring A and/or the group R1 is a substituent containing a heterocyclic group having a substituent at the N-position selected from a lower alkoxycarbonyl group and an aryl-lower alkoxycarbonyl group (e.g., fluorenyl-lower alkoxycarbonyl group, phenyl-lower alkoxycarbonyl group, etc.). The removal of these groups is carried out by the same method as in the above Step (g).
Step (i)
The desired compound (I) wherein the group R1 is a mono-lower alkanoylamino group, a di-lower alkanoylamino group or a mono- or di-lower alkanoylamino-substituted aryl group is prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is an amino group or an aryl group being substituted by an amino group with a lower alkanoic aid or a reactive derivative thereof. The lower alkanoic acid includes, for example, an alkanoic acid having 1 to 6 carbon atoms (e.g., formic acid, acetic acid, propionic acid, etc.). The reactive derivative of alkanoic acid is, for example, an acid halide (e.g., acid chloride, acid bromide, etc.), an acid anhydride, or a mixed acid anhydride. When a free lower alkanoic acid is used, the reaction is carried out in the presence of a condensing agent (e.g., 1,3-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, diethylphosphoric cyanide, diphenylphosphoric azide). When a reactive derivative of the above alkanoic acid is used, the reaction is carried out in the presence of an acid acceptor such as an organic base (e.g., triethylamine, pyridine, etc.), an alkali metal hydroxide, an alkali metal hydrogen carbonate, an alkali metal carbonate, etc. In the reaction, the conversion from the compound (I) wherein R1 is an amino group into the corresponding compound (I) wherein R1 is a mono-lower alkanoylamino group is carried out by controlling the amount of the lower alkanoic aid or a reactive derivative thereof in 0.8-1 equivalent to the amount of the starting compound. The conversion from the compound (I) wherein R1 is an amino group into the corresponding compound (I) wherein R1 is a di-lower alkanoylamino group is carried out by controlling the amount of the lower alkanoic aid or a reactive derivative thereof in 2-3 equivalents to the amount of the starting compound. The reaction is carried out at xe2x88x9230-80xc2x0 C., preferably at xe2x88x9220-50xc2x0 C.
Step (j)
The desired compound (I) wherein the group R1 is an amino group being substituted by one or two groups selected from a lower alkyl group and a hydroxy-substituted lower alkyl group is prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is a mono-substituted or unsubstituted amino group with an allylating agent such as a lower alkyl halide (e.g., a lower alkyl chloride, a lower alkyl bromide, a hydroxy-lower alkyl chloride, a hydroxy-lower alkyl bromide, etc.) wherein the alkyl moiety may optionally be substituted by a hydroxy group, or a lower alkyl-lower alkane-sulfonate (e.g., a lower alkyl methanesulfonate, etc.), lower alkyl arylsulfonate (e.g., a lower alkyl p-toluenesulfonate) wherein the alkyl moiety may optionally be substituted by a hydroxy group in the presence or absence of an acid acceptor. The acid acceptor is, for example, an alkali metal hydroxide (e.g., sodium hydroxide), an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate), an alkali metal carbonate (e.g., sodium carbonate), or an organic base (e.g., triethylamine, pyridine, etc.). The desired compound (I) wherein the group R1 is an amino group being substituted by one group selected from a lower alkyl group and a hydroxy-substituted lower alkyl group is prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is unsubstituted amino group with a lower alkyl aldehyde wherein the alkyl moiety may optionally be substituted by a hydroxy group, and subjecting the product to reduction. The reducing agent is preferably sodium cyanoborohydride, sodium borohydride, sodium triacetoxyborohydride, formic acid, etc. The solvent may be, for example, water, acetic acid, tetrahydrofuran, dioxane, chloroform methylene chloride, methanol, ethanol, etc., or a mixture of these solvents. The reaction is carried out at 0-70xc2x0 C., preferably at 5-50xc2x0 C. The compound (a) wherein the group R1 is an amino group being substituted by a lower alkanoyloxy-lower alkyl group is prepared in the same esterification reaction as in the preparation of the compound [I-b] from the compound [I-a], but preferably prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is an amino group being substituted by a hydroxy-substituted lower alkyl group with a lower alkanoic acid in the presence of a condensing agent (e.g., 1,3-dicyclohexylcarbodiimide, ethyl-3-(3-dimethyl-aminopropyl)carbodiimide hydrochloride, diethyl phosphoric cyanide, diphenyl-phosphoric azide, etc.), or with a reactive derivative of the above lower alkanoic acid such as an active ester (e.g., N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 1-hydroxybenzotriazol ester, etc.), a corresponding acid halide, a corresponding mixed acid anhydride, in the presence of an acid acceptor such as an alkali metal hydroxide (e.g., sodium hydroxide, etc.), an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate, etc.), an alkali metal carbonate (e.g., sodium carbonate, etc.), or an organic base (e.g., triethylamine, pyridine, etc.). The reaction is carried out at xe2x88x9230-80xc2x0 C., preferably at xe2x88x9220-50xc2x0 C.
Step (k)
The desired compound (I) wherein the group R1 is an aryl group being substituted by a di-(lower alkylsulfonyl) amino group is prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is an amino-substituted aryl group with a lower alkyl sulfonyl halide (e.g., a lower alkyl sulfonyl chloride, a lower alkyl sulfonyl bromide, etc.) in the presence of the same acid acceptor as those used in the above Step (j), i.e., an organic base (e.g., triethylamine, pyridine, etc.). The reaction is carried out at 0-80xc2x0 C., preferably at 10-60xc2x0 C.
Step (l)
The desired compound (I) wherein the group R1 is a hydroxy-substituted aryl group, or the desired compound (I) wherein the substituent of Ring A and/or Ring B is a hydroxy group is prepared by removing the protecting groups from a corresponding compound of the formula (I) wherein the group R1 is a protected hydroxy-substituted aryl group, or a corresponding compound of the formula (I) wherein the substituent of Ring A and/or Ring B is a protected hydroxy group. The removal of the protecting group is carried out by a conventional method such as acid-treatment, base-treatment, catalytic reduction, etc. which should be selected according to the kinds of the protecting groups to be removed. The reaction is carried out at 0-80xc2x0 C., preferably at 5-50xc2x0 C.
Step (m)
The desired compound (I) wherein the group B2 of Ring B is a hydroxy group is prepared by treating a corresponding compound of the formula (I) wherein B2 is a lower alkoxy group by a conventional method, such as acid-treatment. The reaction is carried out at 10-150xc2x0 C., preferably at 20-120xc2x0 C.
Step (n)
The desired compound (I) wherein the group R1 is an aryl group being substituted by a group selected from a lower alkylsulfinyl group and a lower alkylsulfonyl group is prepared by oxidizing a corresponding compound of the formula (I) wherein R1 is an aryl group being substituted by a lower alkylthio group. The oxidization reaction is carried out by using an oxidizing agent. The oxidizing agent is, for example, a peroxide compound such as 3-chloroperbenzoic acid, peracetic acid, hydrogen peroxide, trifluoroperacetic acid, etc., sodium periodate, osmium tetraoxide, sodium bromite, etc. When an oxidizing agent is used in an amount of 0.8-1 equivalent to 1 equivalent of the starting compound, there is obtained the compound of the formula (I) wherein R1 is an aryl group being substituted by a lower alkylsulfinyl group. When an oxidizing agent is used in an amount of 2-3 equivalents to 1 equivalent of the starting compound, there is obtained the compound of the formula (I) wherein R1 is an aryl group being substituted by a lower alkylsulfonyl group. The reaction is carried out at xe2x88x9210-60xc2x0 C., preferably at 5-40xc2x0 C.
Step (o)
The desired compound (I) wherein the group R1 is a heterocyclic group being substituted by one or two oxo groups (e.g., a thiomorpholino group being substituted by one or two oxo groups) is prepared by treating a corresponding compound of the formula (I) wherein R1 is a heterocyclic group in the same manner as in the above Step (n).
Step (p)
The desired compound (I) wherein the group R1 is an aryl group being substituted by a mono- or di-lower alkylamino group, or a lower alkyl group being substituted by a mono- or di-lower alkylamino group is prepared in the same manner as in the above Step (j) but is prepared by reacting a corresponding compound of the formula (I) wherein R1 is an amino-substituted aryl group or an amino-substituted lower alkyl group in the presence or absence of an acid acceptor with a lower alkyl halide (e.g., a lower alkyl chloride, a lower alkyl bromide, etc.). The acid acceptor is, for example, an alkali metal hydroxide, an alkali metal hydrogen carbonate, an alkali metal carbonate, an organic base (e.g., triethylamine, pyridine, etc.). When an alkylating agent is used in an amount of 0.8-1 equivalent to 1 equivalent of the starting compound, there is obtained the compound (I) wherein the group R1 is an aryl group (or a lower alkyl group) substituted by a mono-lower alkylamino group. When an alkylating agent is used in an amount of 2-3 equivalents to 1 equivalent of the starting compound, there is obtained the compound (I) wherein the group R1 is an aryl group (or a lower alkyl group) being substituted by a di-lower alkylamino group. The reaction is carried out at 0-60xc2x0 C., preferably at 5-40xc2x0 C. The desired compound (I) wherein the group R1 is an aryl group being substituted by an amino group being substituted by a lower alkyl group and a protecting group for amino group is prepared by treating a corresponding compound of the formula (I) wherein R1 is an aryl group substituted by an amino group being substituted by one protecting group for amino group in the same manner as above. Moreover, the compound (I) wherein the group R1 is a mono-lower alkylamino-substituted aryl group is obtained by removing a protecting group from a corresponding compound of the formula (I) wherein R1 is an aryl group being substituted by an amino group being substituted by a lower alkyl group and a protecting group for amino group by a conventional method.
Step (q)
The compound (I) wherein the group R1 is a pyridylcarbonyloxy-lower alkyl group is prepared by reacting a corresponding compound of the formula (I) wherein R1 is a hydroxy-substituted lower alkyl group with a pyridine-carboxylic acid in the presence of a condensing agent (e.g., 1,3-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, diethyl phosphoric cyanide, diphenylphosphoric azide, etc.), or with a reactive derivative of a pyridinecarboxylic acid (e.g., active ester such as N-hydroxysuccinimide ester, N-hydroxyphthalimide ester, 1-hydroxybenzotriazole ester, etc., pyridinecarboxylic halide) in the presence of an acid acceptor such as an alkali metal hydroxide (e.g., sodium hydroxide, etc.), an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate, etc.), an alkali metal carbonate (e.g., sodium carbonate, etc.), an alkali metal hydride (e.g., sodium hydride, etc.), or an organic base (e.g., triethylamine, pyridine, etc.). The reaction is carried out at 0-60xc2x0 C., preferably at 5-40xc2x0 C.
Step (r)
The compound (I) wherein Ring A is a benzene ring being substituted by a tetrazolyl-lower alkoxy group is prepared by reacting a corresponding compound of the formula (I) wherein Ring A is a benzene ring being substituted by a cyano-lower alkoxy group, with a metal azide such as sodium azide, tributyltin azide. The reaction is carried out at 30-120xc2x0 C., preferably at 50-100xc2x0 C.
Step (s)
The compound (I) wherein the substituent of Ring A is a group containing a heterocyclic group substituted by an oxo group (e.g., an oxo-substituted pyridyl-substituted lower alkyl group) is prepared by treating a corresponding compound of the formula (I) wherein the substituent of Ring A is a group containing a heterocyclic group with an oxidizing agent (e.g., 3-chloroperbenzoic acid, hydrogen peroxide, peracetic acid, etc.). The reaction is carried out in the same manner as in the above Step (n).
Step (t)
The compound (I) wherein the group R1 is a heterocyclic group (e.g., piperazinyl group) having a hydroxy-lower alkyl group at the N-position is prepared by reacting a corresponding compound of the formula (I) wherein the group R1 is a heterocyclic group with a lower alkyl halide (e.g., hydroxy-lower alkyl chloride, hydroxy-lower alkyl bromide, etc.) wherein the alkyl moiety is substituted by a hydroxy group, in the presence or absence of the same acid acceptor (e.g., an alkali metal carbonate such as sodium carbonate) as those used in the above Step (j). The reaction is carried out at 40-120xc2x0 C., preferably at 50-100xc2x0 C.
The solvent used in the above Steps (a) to (t) may be any one which does not disturb the reaction, for example, dioxane, ethylene glycol dimethyl ether, dimethyl acetamide, dimethylfonnamide, hexamethylphosphoramide, benzene, tetrahydrofuran, toluene, ethyl acetate, a lower alcohol, methylene chloride, chloroform, carbon tetrachloride, 1,3-dimethyl-2-imidazolidinone, acetic acid, diethyl ether, dimethoxyethane, dimethylsulfoxide, water, or a mixture of these solvents.
The starting compound (II) is prepared, for example, by reacting a benzoylbenzoic acid compound of the formula (i):
wherein the symbols are the same as defined above, with a malonic acid compound of the formula (ii):
X2xe2x80x94CH(COOZ1)2
wherein X2 is a leaving group, and Z1 is a protecting group for carboxyl group, in the presence of a base,
removing the protecting group from the product to give a compound of the formula (iii):
wherein the symbols are the same as defined above;
subjecting the compound (iii) to decarboxylation reaction and dehydration reaction in the presence or absence of an acid to give a compound of the formula (iv):
wherein the symbols are the same as defined above;
if necessary, followed by converting the 3-carboxyl group of the compound (iv) into the substituent R2 by esterification or amidation by a conventional method.
The starting compound (V) is prepared, for example, by condensing the compound of the above formula (i) with a compound of the formula (v):
wherein the symbols are the same as defined above, in the same manner as in the condensation reaction between the compound (I-a) and the amine compound (VIII).
The benzoyl benzoic acid compound (i) is prepared by a conventional method, for example, by treating a benzaldehyde compound of the formula (vi):
wherein Ring A is the same as defined above, with a halogen (bromide, etc.),
reacting the resulting o-halogeno benzaldehyde compound with an acetalization agent, for example, with a lower alkyl orthoformate (e.g., methyl orthoformate, etc.), in the presence of an acidic catalyst (e.g., a strong acidic resin, etc.) to protect the formyl group,
reacting the product with an aldehyde compound of the formula (vii):
wherein Ring B is the same as defined above, oxidizing the product, i.e., treating with an oxidizing agent such as manganese dioxide, etc., to give a compound of the formula (viii):
wherein the symbols are the same as defined above,
subjecting the compound (viii) to deacetalization by treating with an acid (e.g., hydrochloric acid, trifluoroacetic acid, a strong acidic resin),
followed by treating with an oxidizing agent (e.g., sodium chlorite).
Besides, in the preparation of the compound (i) as mentioned above benzoic acid compound of the following formula (ix) is also used instead compound (vii).
wherein W is a di-lower alkyl-substituted carbamoyl group, a lower alkoxycarbonyl group, or a carboxyl group forming a salt with an alkali metal (e.g., sodium, potassium, etc.), and Ring B is the same as defined above.
Moreover, the starting compound (i) of the present invention is prepared by reacting a compound of the formula (x):
wherein a group of the formula: is a heterocyclic group which may optionally be substituted by a lower alkyl etc., and Ring A is the same as defined above, with the compound (vii) in the presence of a base (e.g., n-butyl lithium, etc.) to give a compound of the formula (xi):
wherein the symbols are the same as defined above;
heating the compound (xi) in the presence of an acid (e.g., an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, etc.) to give a compound of the formula (xii):
wherein the symbols are the same as defined above;
subjecting the compound (vii) to hydrolysis with a base (e.g., an alkali metal hydroxide such as potassium hydroxide, sodium hydroxide, etc.);
then, followed by subjecting the product to oxidation.
The compound (xii) is prepared by reacting a compound of the formula (xiii):
wherein Y is a mono- or di-lower alkylamino group, and Ring A is the same as defined above, with the compound (vii) in the presence of a base (e.g., sec-butyl lithium, etc.) to give a compound of the formula (xiv):
where the symbols are the same as defined above, followed by heating the compound (xiv) in the presence of an acid such as an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, etc.).
Throughout the present description and the claims, the xe2x80x9calkyl groupxe2x80x9d means an alkyl group having 1 to 16 carbon atoms, and preferably a straight chain or branched chain alkyl group having 1 to 8 carbon atoms. The xe2x80x9clower all groupxe2x80x9d, the xe2x80x9clower alkoxy groupxe2x80x9d and the xe2x80x9clower alkylene groupxe2x80x9d mean ones having 1 to 6 carbon atoms, respectively, and preferably ones having 1 to 4 carbon atoms. The xe2x80x9clower alkenyl groupxe2x80x9d and the xe2x80x9clower alkynyl groupxe2x80x9d mean ones having 2 to 7 carbon atoms, respectively, and preferably a straight chain or branched chain one having 2 to 5 carbon atoms. The xe2x80x9clower alkylenedioxy groupxe2x80x9d and the xe2x80x9calkanoyl groupxe2x80x9d mean ones having 1 to 7 carbon atoms, respectively, and preferably a straight chain or branched chain one having 1 to 5 carbon atoms. The xe2x80x9ccyclo-lower alkyl groupxe2x80x9d means cycloalkyl groups having 3 to 8 carbon atoms, preferably ones having 3 to 6 carbon atoms.

Method of inhibiting neoplastic cells with isoquinoline derivatives


The present invention provides methionine aminopeptidases (MetAPs) with a broad substrate range, particularly those capable of removing the N-terminal Met from bulky or acidic penultimate residues. In preferred embodiments, these MetAPs have mutations at the 233, 206 and/or 168 positions of SEQ ID NO:1. Preferably, amino acids at these residues are substituted with glycine or threonine. Also provided are cells comprising the MetAPs, DNA encoding the MetAPs, and methods of using the MetAPs.

Removal of the translation initiator N-formyl-methionine or methionine from a recombinant protein is often critical for its function and stability. For example, it has been shown that such removal is essential for human hemoglobin, interleukin-2, growth hormones or frog ribonucleases (Adachi et al., 2000; Endo et al., 2001; Busby et al., 1987; Liao et al., 2003; Boix et al., 1996; Varshavsky, 1996). For the preparation of proteins with innate N-terminus, various attempts have been made to remove the N-terminal Met. First, cyanogen bromide (CNBr) was used to cleave Met under extreme acidic conditions, but the method is limited to proteins without any internal Met residue (Boix et al., 1996). Second, a protease-specific oligopeptide was introduced in front of a target protein, which was then removed in vitro by the respective protease, e.g., factor Xa, enterokinase or cathepsin C (Belagaje et al., 1997). Third, the N-terminal Met of a protein was removed in vitro by the aminopeptidase of Aeromonas proteolytica (Notomista et al., 1999; Shapiro et al., 1988). Fourth, a signal peptide was introduced in front of target protein and processed in vivo during secretion, but the yield was low (1˜5 mg per liter of culture) (Huang et al., 1998).
Recently, methionine aminopeptidases (MetAPs) were employed to remove the N-terminal Met in E. coli and yeast. There are two types of MetAP, MetAP I and MetAP II, in eukaryotes. Only MetAP I exists in eubacteria; MetAP II is present in archaea (Li et al., 1995; Tahirov et al., 1998). It has been demonstrated that the genes for these MetAPs are essential for the growth of prokaryotes and eukaryotes (Li et al., 1995; Chang et al., 1989). However, the efficiency of Met removal is limited by the size of the side chain (radius<3.68 Å) of the penultimate residue, the residue next to the N-terminal Met (Ben-Bassat et al., 1987; Hirel et al., 1989; Hwang et al., 1999; Chen et al., 2002). Thus, Met removal for proteins with larger or bulky penultimate residues is inefficient. It is also difficult to remove the N-terminal Met when the penultimate residue is charged, either acidic (Glu or Asp) or basic (Lys, Arg or His).
From the structure of E. coli MetAP and bestatin-based inhibitor complex, it was discovered that four residues (Tyr168, Gln233, Met206 and Glu204) reside in the substrate binding pocket (Lowther et al., 1999; Lowther et al., 2000). The Met329 and Gln356 residues of yeast MetAP, corresponding to Met206 and Gln233 of E. coli MetAP, were replaced with Ala. When tested in vitro, these purified MetAP I variants (M329A, Q356A) had significantly increased catalytic activities for oligopeptides with slightly larger penultimate residues, such as Asn, His and Met, but not with bulky or acidic penultimate residues (Roderick et al., 1993; Walker et al., 1999). Therefore, the need exists for a method to remove N-terminal Met that is more universally applicable, particularly a method that can remove the N-terminal Met from a bulky or acidic penultimate residue.

Removal of N-terminal methionine from proteins by engineered methionine aminopeptidase