HK1023510A - A combined preparation comprising 10, 11-methanodibenzosuberane derivatives - Google Patents

Description

A compound product containing 10, 11-methylene dibenzosuberane derivatives

This application is a divisional application filed as application No. 94191807.6, filed as 19940413, entitled "10, 11-methylenedibenzosuberane derivatives for use as chemosensitizers".

The present invention relates to pharmaceutically active substances. These substances are used for the treatment of cancer, in particular to enhance the efficacy of existing cancer chemotherapeutic drugs and to treat multidrug resistance. More specifically, the present invention relates to a series of 10, 11-methylene dibenzosuberane derivatives. The invention also relates to pharmaceutical formulations and methods of chemosensitization, for example in the treatment of cancer, including reversal of multidrug resistance, and to the use and methods of preparation of novel compounds in the preparation of pharmaceutical compositions containing these substances.

One of the problems faced with cancer chemotherapy is the development of resistance to the treatment regimen. Some tumors initially respond well to certain drugs, but resistance to these drugs often occurs. This disease state, known as multidrug resistance, is discussed in detail in Kuzmich and Tew, section VII, in particular, "The Multi-ug-resistant Phototype (MDR)," Medical Research Reviews, Vol.11, No.2,185-217, in particular 208-213 (1991); and Georges, Sharom and Ling, "Multi drug Resistance and Chemosentizati-on: Therapeutic injections for Cancer Chemotherapy," Advances in Pharmacology, Vol.21,185-220 (1990).

Some active substances, called chemosensitizers or potentiators, have been used as resistance-modifying agents to overcome multi-drug resistance, but suffer from a number of disadvantages. These include, for example, verapamil (a calcium channel blocker which lowers blood pressure and has been found to be effective in vitro for the treatment of drug-resistant malaria), steroids, trifluoro-10- (gamma-methylpiperazinopropyl) -phenothiazine (a CNS agent), vinpocetine, and reserpine (a-2 blocker with CNS properties). Thus, there remains a need for active substances for use in therapy, i.e. reversing, inhibiting and/or preventing multidrug resistance, and preferably with little or no side effects.

The chemosensitizers interact with P-glycoprotein, a drug efflux pump found on cell membranes, particularly those of tumor cells with multidrug resistance, gastrointestinal cells and endothelial cells that form the blood-brain barrier, and by blocking this pump, the chemosensitizers can inhibit the efflux of cancer chemotherapeutic drugs from tumor cells, thereby enhancing the penetration of nutrients or active substances into the gastrointestinal tract and the penetration of active substances through the blood-brain barrier.

Fukazawa et al, in U.S. patent No.5,112,817, disclose quinoline derivatives useful as potentiators of anticancer drugs for the treatment of multidrug resistance. One of the initially more promising active substances disclosed therein is MS-073, which has the following structure:

however, MS-703, which is highly active in vitro, has low bioavailability when administered orally and is unstable in solution. Other compounds of this series, such as the diphenylmethylcarbonyl derivative MS-209, while having good stability and oral bioavailability, require higher effective doses to be taken. Therefore, there is a problem in providing a potentiator of an anticancer drug, which has both the activity of MS-073 and good oral bioavailability and stability. The present invention aims to solve this problem.

One aspect of the present invention is directed to 10, 11-methylene dibenzosuberane derivatives, and pharmaceutically acceptable salts thereof, of the formula:formula i wherein:

a is-CH₂-CH₂-,-CH₂-CHR^a-CH₂-, or-CH₂-CHR^a-CHR^b-CH₂-, wherein R^aOr R^bOne is H, DH or lower acyloxy and the other is H;

R¹is H, F, Cl or Br;

R²is H, F, Cl or Br; and is

R³Is substituted by F, Cl, Br, CF₃、CN、NO₂Or OCHF₂Optionally substituted heteroaryl or phenyl.

The present invention preferably relates to certain compounds of formula I, especially the single isomers thereof, including especially those wherein A is-CH₂-CHR^a-CH₂-、R^aIs OH, R¹Is F, R²Is F and R³Are quinolinyl compounds, the most preferred being the (2R) -trans isomer.

In another aspect, the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, which pharmaceutical composition will also include a pharmaceutically acceptable excipient.

In another aspect, the invention relates to a method of treatment by administering to a mammal in need thereof, a compound of formula I or a pharmaceutically acceptable salt thereof, in a therapeutically effective amount to enhance the efficacy of a co-administered cancer chemotherapeutic agent, or by using a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition. The cancer chemotherapeutic drugs mentioned above are antimetabolites such as 6-mercaptopurine, 5-fluorouracil, cytarabine and metabolites or derivatives of these substances. Other cancer chemotherapeutic agents are the antidiolates such as methotrexate or derivatives of certain natural products, for example, derivatives of vinca alkaloids, vinblastine, vincristine and colchicine; adriamycin, daunorubicin, doxorubicin, epipodophyllotoxin thiophene polyglycoside or etoposide. Such cancer chemotherapeutic agents also include platinum anti-cancer agents, such as cisplatin and carboplatin. In addition, the medicament of the present invention may be administered together with such drugs as cyclophosphamide, busulfone, procarbazine, dacarbazine, carmustine, lomustine, mechlorothiamine, chlorram-bucil, hydroxyurea, melphalan, mitotone, paclitaxel and spirogermanium. Multidrug resistance is most commonly seen in vinca alkaloids, anthracyclines, doxorubicin, and doxorubicin; and etoposide and epipodophyllotoxin thiopheneglycoside; less commonly found in antimetabolites and other chemotherapeutic drugs.

In a further aspect, the invention relates to a method of treating drug resistance in a mammal by administering to a mammal in need thereof a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. One embodiment of this aspect may detail a method of treating drug-resistant malaria. In a preferred embodiment, the method of treating multi-drug resistance in a mammal having significant clinical resistance to a cancer chemotherapeutic agent is by co-administering a resistance-modifying dose of a compound of formula I or a salt thereof with a therapeutically effective amount of a cancer chemotherapeutic agent for which resistance is already apparent.

The invention also relates to a method for enhancing the bioavailability of a pharmaceutically active substance, which comprises administering to a mammal in need of such treatment an amount of a compound of formula I or a salt thereof which is capable of increasing the penetration of the active substance through the blood-brain barrier or the gastrointestinal tract.

The invention also relates to a method for preparing the compound shown in the formula I.

The following definitions are used to describe and define the meaning and scope of various terms in this specification.

"alkyl" refers to a saturated monovalent radical containing only carbon and hydrogen, which may be cyclic, branched, or straight-chain. This term may be further described by the following radicals such as methyl, ethyl, t-butyl, pentyl, neopentyl, heptyl and adamantyl.

"lower alkyl" refers to a cyclic, branched, or straight chain monovalent alkyl radical having one to six carbon atoms. This term may be further described by the following radicals such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl (or 2-methylpropyl), cyclopropylmethyl, isopentyl, n-pentyl and hexyl.

"alkylene" refers to a saturated divalent radical containing only carbon and hydrogen, and may be branched or straight-chain. This term can be further described by the following radicals, such as methylene, ethylene, n-propylene, t-butylene, isopentenylene (i-pentylene), and n-heptylene (n-heptylene).

"lower alkylene" refers to a divalent alkyl radical of 1 to 6 carbon atoms. This term can be further described by the following radicals, such as methylene, ethylene (ethylene), n-propylene (n-propylene), i-propylene (i-propylene), n-butylene (n-butylene), t-butylene (t-butylene), i-butylene (i-butylene) (or 2-methylpropylene), iso-pentylene (isoamylene), pentylene (pentylene), and n-hexylene (n-hexylene).

"lower acyloxy" refers to the group-O- (CO) -R 'where R' is lower alkyl.

"aryl" means a monovalent unsaturated aromatic carbocyclic radical, in particular a radical of 6 to 16 carbon atoms having one monocyclic (e.g. phenyl) or two condensed rings (e.g. naphthyl), which may be optionally mono-, di-or trisubstituted independently by fluorine, chlorine, bromine, trifluoromethyl, cyano, nitro and/or trifluoromethoxy.

"heteroaryl" means a monovalent unsaturated aromatic heterocyclic radical, particularly a radical containing from 2 to 12 carbon atoms in the ring of at least one heteroatom, preferably 1-3 heteroatoms such as N, O or S, and is typically monocyclic, such as pyridyl or two condensed rings such as quinolinyl, benzofuranyl, benzofurazanyl.

"halogen" refers to fluorine, bromine, chlorine and iodine.

"optional" or "optionally" means that the stated circumstance may or may not occur thereafter, i.e., the description includes instances where the stated circumstance occurs and instances where it does not.

The "pharmaceutically acceptable salt" may be any salt of an inorganic or organic acid. "pharmaceutically acceptable anion" refers to the anion in such an addition salt. The salts or anions are selected only to be those which are not physiologically active.

The anion is derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid (giving sulfates and sulfites), nitric acid, phosphoric acid, and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, salicylic acid, p-toluenesulfonic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, lactic acid, o- (4-hydroxyphenoxy) benzoic acid, 1, 2-ethyldisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2 ] oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4' -methylenebis- (3-hydroxy-2-naphthalenemethyl) acid, acetic acid, benzoic, 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, lauryl sulfuric acid, glucuronic acid, glutamic acid, 3-hydroxy-2-naphthoic acid, stearic acid, muconic acid, and the like.

"treatment" refers to the treatment of a disease in a mammal, including:

arresting the disease, meaning that the clinical symptoms of the disease do not develop;

(ii) inhibiting disease, meaning arresting the development of clinical symptoms; and/or

(iii) relief of disease, meaning alleviation of clinical symptoms.

An "effective amount" refers to a dosage sufficient to provide treatment of the disease state being treated, depending on the patient, the condition, and the condition being treated.

By "co-administered" is meant that more than one active is administered as part of the same treatment regimen, whether taken at the same time or at different times.

"Structure of formula I" refers to the general formula of the compounds of the present invention. The chemical bonds represented by the curves in formula II refer to non-specific stereochemistry, for example the 5-position of dibenzosuberane, i.e.the piperazinyl group is attached to the carbon atom at this position.

"isomerization" refers to compounds having the same atomic mass and number of atoms but differing in one or more physical or chemical properties.

"stereoisomer" refers to one of two compounds having the same molecular weight, the same chemical composition and composition, but differing in the spatial arrangement of its atoms. That is, certain chemically identical moieties have different orientations in space. Thus, the substance has the ability to rotate the plane of polarized light when purified. However, some pure stereoisomers are difficult to detect with existing instruments because of their weak optical activity.

"optical isomers" describe a type of stereoisomerism characterized by rotation of the plane of polarized light by pure or in solution isomers. In many instances, it is due to the attachment of four different atoms or groups to at least one carbon atom in the molecule. These isomers may be described as d-, l-or d, l-; or D-, L-or D, L-; and may also be described as (R) -, (S) -or (R, S) -, with the different methods of description depending on the nomenclature used.

The compounds of formula I exist in two isomeric configurations, determined by the relationship of the 10, 11-methylene and 5-piperazinyl substituents in dibenzoheptane (see, for example, the structure represented by formula II in the nomenclature below). When the 10, 11-methylene and 5-piperazinyl substituents are in the same orientation relative to the dibenzoheptane (e.g., both up or both down), the isomeric form is referred to as "cis". When the 10, 11-methylene and 5-piperazinyl substituents are in opposite directions (e.g., one above, one below) relative to the dibenzoheptane, the isomers are referred to as "trans".

Certain compounds of formula I have an asymmetric center in the group designated "A", wherein R^aOr R^bIs not hydrogen. These compounds may exist in two stereoisomeric forms, referred to as (+) and (-), or as (R) -and (S) -, or as a mixture of two isomers. The nomenclature of (R) -and (S) -is used in this specification.

Although specific stereoisomers are disclosed and named, the present invention includes individual specific stereoisomers as well as mixtures and racemates thereof. Name of

The compounds of formula I are named and numbered with reference to the compounds of formula II below.Formula II

For example, wherein R¹And R²Is chlorine, R^aIs hydroxy and phenyl (R in formula I)³) The compound in which the 3-position is substituted with a nitro group may be named (3R, S) -trans, cis-1- {4- [4- (10, 11-dichloromethyl dibenzosuberyl-5-yl) piperazin-1-yl-3-hydroxybutoxy } -3-nitrobenzene.

Wherein R is¹And R²Is hydrogen, R^bIs acetoxy (in the form of the downward isomer viewed from the paper) and phenyl (R in formula I)³) Is substituted with a trifluoromethyl group, the bond linking the 4-position of the piperazine to the 5-position of the benzocycloheptane is in the upward isomeric form when viewed from the plane of the paper, and the compound is designated (2S) -cis-1- [4- [4- (10, 11-methylenedibenzocyclohept-5-yl) piperazin-1-yl]-2-acetoxybutoxy } -5-trifluorotoluene.

Preferred compounds of the invention are represented by formula iii:formula III

The compound is wherein R¹And R²Is F, A is (2R) -hydroxypropyl and R³The compound of formula I, which is quinolinyl with oxygen at the 5-position, is named (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl]-2-hydroxypropoxy } quinoline. In addition, the chemical abstracts nomenclature system names the compound of formula III as 1- (4-trans (1, 1-difluoro-1 a,10 b-dihydrodibenzo [ a.e ]]Cyclopropyl [ c ]]Cyclohepten-6-yl) piperazin-1-yl) - (2R) -3- (5-quinolinyloxy) -2-propanol (numbering in formula III does not correspond to the nomenclature system of the chemical abstracts). While any nomenclature is sufficient to describe the compounds of the invention, the former system is used in the description of the invention.

"solvent", "inert organic solvent" or "inert solvent" means that the solvent is inert under the reaction conditions described therein and includes, for example, benzene, toluene, acetonitrile, Tetrahydrofuran (THF), Dimethylformamide (DMF), chloroform, dichloromethane, diethyl ether, methanol, pyridine, and the like). The solvent used in the reaction of the present invention is an inert organic solvent unless otherwise specified.

The term "suitable amount" means an amount added sufficient to achieve the stated effect, e.g., to provide a solution or a desired volume (i.e., 100%).

Unless otherwise indicated, the reactions described herein are generally carried out at 5 ℃ to 100 ℃ under normal pressure (preferably 10 ℃ to 50 ℃; particularly preferably at room or ambient temperature, e.g., 20 ℃). There are some reactions in which the temperature ranges used in the chemical reaction are above or below these temperature ranges. In addition, unless otherwise specified, the time and conditions of the reaction are generally about 1 to about 10 hours (preferably about 5 hours) at 5 to 100 ℃ (preferably about 10 to 50 ℃, particularly preferably 20 ℃) under normal pressure. The parameters given in the examples are all precise and not approximate.

The isolation and purification of the compounds and intermediates described therein may be carried out by an appropriate method such as filtration, extraction, recrystallization, column chromatography, thin layer chromatography or thick layer chromatography or a combination of methods, as required. Specific reference may be made to the following examples for suitable methods of isolation, but other equivalent methods of isolation or purification may be used. Synthesis of Compounds of formula I

The compounds of formula I can be prepared by the methods described in U.S. Pat. No.5,112,817 and the references therein, by replacing dibenzosuberone with an optionally substituted dibenzosuberone. The former can be used by Ciganek, et al, "Imine antibodies of Tricyclic anti-depressor-ssant," J.Med.chem.,1981,24, 336-4; or Coyne and Cusic, "amino alkyldibenzozo [ a, e ] cyclopropa [ c ] cyclohextene Deriv-coatings, A Series of content antioxidants," J.Med.Chem.1974, Vol.17, No.1,72-75, and the like (and references therein). Various synthetic methods for compounds of formula I are described below, with reference to schemes 1,2 and 3.

The compound of formula 4 below 1- [10, 11-methylenedibenzocycloheptan-5-yl]The piperazines may be optionally substituted at the 10 and 11 positions, which play a key role in the synthesis of the compounds of formula I. The key intermediate and the compound containing the group A-O-R³Coupling or condensing the reagents to obtain or convert the compound of formula I; or with an A-epoxy group or a derivative thereof and A' -O-R³Condensing the group with a reagent to obtain or convert the compound of formula I. This conversion includes, for example, acylation of the hydroxyl group in the radical A 'to an acyloxy group, or removal of the acyloxy group in the radical A', or use of R³-OH to convert the group A-epoxide to a compound of formula I. In this transformation, when R is³when-OH is added to the group A-epoxy, the epoxy ring is opened, with-OR³The adjacent carbon of the connecting carbon forms a hydroxyl group. Brief description of the reaction scheme

Scheme 1 illustrates the synthesis of compounds of formula I wherein R^aIs H or OH.

Scheme 2 illustrates the synthesis of compounds of formula 8 and formula 9, which are useful as precursors for the synthesis of compounds of formula I (see scheme 1, step 5).

Scheme 3 illustrates the synthesis of compounds of formula I wherein R^bIs OH.

Substituent A, R used in the reaction scheme¹、R²、R³、R^aAnd R^bAnd have the same meaning as in the preceding summary. The substituent X represents a halogen atom; n is 1 or 2; m is 1,2, 3 or 4. Starting materials

The compound 5H-dibenzo [ a, d ] cyclohepten-5-one (also known as dibenzo [ a, d ] -5H-cyclohepten-5-one or dibenzosuberone) is commercially available, for example, from Aldrich chemical company. Other reactants, like propylene oxide bromide and 1-bromo-3, 4-butylene oxide are also commercially available or can be readily prepared by the skilled person using conventional synthetic methods.

Reaction scheme 1Preparation of the Compound of formula 1

Acetates (e.g. sodium chlorodifluoroacetate, methyl trichloroacetate, ethyl trifluoroacetate, according to the desired substituent R¹And R²Defined) is added to a solution of dibenzosuberone (e.g. in diglyme) in a solvent (e.g. diglyme, benzene or petroleum ether) over a period of 4 to 8 hours (preferably 6 hours) with stirring and nitrogen protection, the reaction temperature being maintained at 160-165 ℃. (other reaction temperatures may be used, depending on the reactants used, as described by Ciganek, et al and Coyne with Cusic et al.) when the reaction mixture reaches room temperature, it is poured into water and extracted (e.g., with ether). The desired 10, 11-substituted methylene dibenzosuberone can be isolated and purified by conventional methods, for example, the organic phase (e.g., benzene) can be washed with water and dried (e.g., with Na)₂SO₄) Evaporated and the residue recrystallized (e.g. from ethanol or optionally once more, e.g. from acetone/hexane).

Alternatively, R¹And R²Compounds of formula 2 which are different (e.g., H and Cl, respectively) can be prepared by the method of j.med.chem.vol.17, 72(1974), which is incorporated herein by reference. R¹And R²Compounds of formula 2 which are both hydrogen can be used as Coyne and Cusic "Aminoalkyl di-benzol [ a, e]cyclopropa[c]Cycloheptene derivatives. A series of patent antibodies ", J.Med.chem.,1974, Vol 17, No.1,72-75, which is incorporated herein by reference. Preparation of the Compound of formula 2

A solution of 10,11- (optionally substituted) -methylenedibenzosuberone in a solvent (e.g., THF/methanol) is cooled (e.g., in an ice bath) and a reducing agent (e.g., sodium borohydride) is added in portions. The reaction mixture was allowed to return to room temperature and stirred for 1 to 5 hours (preferably 2 hours), then poured into water. The product is isolated (e.g., filtered) and purified by conventional means (e.g., water washing and drying) to provide the corresponding 10,11- (optionally substituted) -methylenedibenzosubenol. Preparation of compound of formula 3 wherein R is formyl

A solution of 10,11- (optionally substituted) -methylene-dibenzosubenol in a solvent (e.g. dioxane) is cooled (e.g. in an ice bath) and then halogenated [ e.g. thionyl chloride is added dropwise, maintaining the temperature (40 to 70 ℃, preferably 50 ℃) for 2 to 5 hours (preferably 4 hours)]. The reaction mixture was evaporated to dryness to give a mixture of cis and trans isomers of the corresponding 5-halo-10, 11- (optionally substituted) -methylenedibenzosuberane. This halogenated cycloheptane is dissolved without further purification (e.g. in acetonitrile) and piperazine is introduced by nucleophilic substitution of the halide [ e.g. under stirring, preferably under dry N ]₂Adding 1-piperazinecarboxaldehyde under atmosphere and increasing temperature (such as 100 deg.C), and taking 10 to 30 hr (preferably 20 hr)]. The reaction mixture is evaporated to dryness and the residue is purified by conventional means [ e.g.by subjecting the residue to NaHCO₃Partitioning between the solution and ethyl acetate, washing the organic phase with water, and drying (e.g. with K)₂CO₃) And evaporate]Isolation and purification of the desired 1- [10,11- (optionally substituted) -methylene-dibenzocycloheptan-5-yl radical]-4-formylpiperazine. Each of which isThe formula and trans isomers can be separated by flash silica gel chromatography (30% acetone/hexane). Preparation of the Compound of formula 4

1- [10,11- (optionally substituted) methylenedibenzocycloheptan-5-yl group]A solution of-4-formyl-piperazine and potassium hydroxide in a solvent (e.g., 9: 1 ethanol/water) is refluxed for 0.5 to 2 hours (preferably 1 hour), and then cooled. The cooled reaction mixture is concentrated, diluted with water, extracted (e.g. with ethyl acetate) and dried (e.g. with K)₂CO₃) The organic phase is evaporated off to give the corresponding 1- [10,11- (optionally substituted) methylenedibenzosuberane-5-yl radical]Piperazine. Preparation of Compounds of formula I

A compound of formula 8 [ e.g., 1- (aryloxy or heterocyclyloxy) -2, 3-epoxypropane or 1- (aryloxy or heterocyclyloxy) -3, 4-epoxybutane ] or an aryloxy or heterocyclyloxy-alkyl halide of formula 9 and 1- [10,11- (optionally substituted) methylenedibenzocyclohept-5-yl ] piperazine are refluxed in a solvent (e.g., isopropanol) for 10 to 30 hours (preferably 20 hours). The desired 10, 11-methylenedibenzosuberane derivative of the formula I is isolated and purified by customary methods [ e.g.by evaporation to dryness, chromatography on silica gel (e.g.with 70: 30: 1 ethyl acetate/cyclohexane/triethylamine) ].

Reaction scheme 2Preparation of the Compound of formula 8

An aromatic or heterocyclic aromatic alcohol (e.g., benzofurazan-4-ol, quinolin-5-ol, or 2-nitrophenol) is dissolved in a solvent (e.g., acetonitrile, THF, or dimethylformamide) and treated with a slight excess of a strong base (e.g., sodium hydride or potassium tert-butoxide). The mixture is heated (e.g. at 50 ℃) for 10 minutes to 2 hours (preferably 30 minutes). A compound of formula 6 (e.g., 1-chloro-2, 3-epoxybutane, 1-bromo-2, 3-epoxybutane, epibromopropane, epichlorohydrin, or their p-toluenesulfonyl or methanesulfonyl derivatives) is added and the mixture is heated (e.g., at 60 ℃ for 1 to 5 hours; preferably for 2 hours). The reaction mixture is poured into water and extracted (e.g., with ethyl acetate). The organic phase is washed with water, Na₂SO₄Drying and evaporating to obtain the corresponding1- (Aryloxy or heterocyclyloxy) -2, 3-epoxypropane or 1- (Aryloxy or heterocyclyloxy) -3, 4-epoxybutane, which can be prepared by conventional methods [ e.g., silica gel chromatography (50% acetic acid/hexane)]And (5) separating and purifying.

Compounds of formula 8, such as 1- (5-quinolinyloxy) -2, 3-cyclopropane, can also be prepared by the method of drug, Desi-gn and Discovery, Vol 9,69(1992), which is incorporated herein by reference. Preparation of the Compound of formula 9

As shown in scheme 2, aromatic or heteroaromatic anions of formula 5 are reacted with dihaloalkyl compounds of formula 7, such as 1-bromo-2-chloroethane, 1-bromo-3-chloropropane or 1-bromo-4-chlorobutane, in a solvent (such as acetone, THF or DMF) at temperatures ranging from room temperature to the boiling point of the solvent to form the corresponding haloalkoxyaryl (or heteroaromatic) compounds of formula 9. This synthesis is described in U.S. patent No.5,112,817, which is incorporated herein by reference.

Reaction scheme 3Preparation of Compounds of formula 10

As shown in scheme 3, step 1, Compound 1- [10,11- (optionally substituted) -methylenedibenzocycloheptan-5-yl of formula 4]Piperazine is reacted with a compound of formula 6 (e.g., a compound of scheme 3 with n = 2; other compounds of formula 6 give the corresponding products), under conditions relevant to the preparation of the compound of formula 8 above (scheme 2), to give the compound of formula 10, 1- [10,11- (optionally substituted) methylenedibenzocycloheptan-5-yl]-4- (3, 4-epoxybutyl) piperazine. R^bPreparation of compounds of formula I which are OH

As shown in scheme 3, step 2, compounds of formula 10, 1- [10,11- (optionally substituted) -methylenedibenzocycloheptan-5-yl]Reaction of (E) -4- (3, 4-epoxybutyl) piperazine with an aromatic or heterocyclic aromatic alcohol (e.g., benzofurazan-4-ol, quinolin-5-ol, or 2-nitrophenol) under the conditions described above in connection with the preparation of the compound of formula I (scheme 1) gives R^bThe corresponding compound of formula I being OH. R^aOr R^bPreparation of compounds of formula I which are lower acyloxy

R^aOr R^bCompounds of formula I which are low acyloxy may be prepared as described in U.S. Pat. No.5,112,817 to which reference is made^aOr R^bStarting from the corresponding compound which is OH (prepared as described above). For example, R^aOr R^bThe compound of formula i, which is OH, is reacted with an acid chloride to produce the corresponding acyloxy compound. Preparation of salts of the Compounds of formula I

The compounds of formula I can be converted into the corresponding acid addition salts. The conversion is accomplished by treatment with a stoichiometrically equivalent amount of a suitable acid, such as hydrochloric acid (e.g., 3 molar equivalents of trihydrochloride in the case of compounds of formula I containing 3 basic nitrogen atoms). If R is³For phenyl, the compound of formula i has only two basic nitrogens and will absorb two equivalents of the acid to form an acid addition salt. If the substituent R is³Including two basic nitrogen atoms, the base of formula i will absorb four equivalents of acid. Preferred acid addition salts of the present invention comprise 2 or 3 equivalents of the acid. Most preferred are acid addition salts with 3 equivalents of acid. In a typical salt-forming step of the invention, the free base is dissolved in a polar organic solvent, such as methanol or ethanol, and the acid is added to water, methanol or ethanol. The temperature is maintained between 0 ℃ and 50 ℃. The corresponding salt is precipitated simultaneously, or precipitated out with a weakly polar solvent, or by evaporating the solvent, or cooling the solution.

According to the invention, the acid addition salts of formula I can be decomposed into the corresponding free bases by treatment with an excess of a suitable base, such as ammonia or sodium bicarbonate, in aqueous solution and at a temperature in the range from 0 ℃ to 50 ℃. The free base can be isolated by conventional methods, such as extraction with an organic solvent. Obviously, the stoichiometric excess must take into account the number of equivalents of acid bound by the base of formula i.

Preferred free bases of formula I include 2 or 3 basic nitrogen atoms. Bases with 3 nitrogen atoms are most preferred.

Preferred method and final step

1- (aryloxy or heteroaryloxy) -2, 3-epoxypropane or 1- (aryloxy or heteroaryloxy) -3, 4-epoxybutane or an aryloxy or heteroaryloxyalkyl halide is combined with 1- [10,11- (optionally substituted) methylenedibenzocycloheptan-5-yl ] piperazine to form the corresponding 10, 11-methylenedibenzocycloheptan derivatives of formula I.

R^aOr R^bThe compound of formula i, which is OH, is reacted with an acid chloride to produce the corresponding acyloxy compound.

Wherein is OH, a compound of formula 10 and an alcohol R₃-OH to form the corresponding 10, 11-methylenedibenzosuberane derivative of formula I.

The compounds of formula I are combined with a pharmaceutically acceptable acid to form the corresponding acid addition salts.

The pharmaceutically acceptable acid addition salts of formula I are reacted with a base to form the corresponding free base of formula I.

Preferred compounds

Preferred compounds are R¹And R²A compound of formula 1 which is fluorine. A compound wherein A is 2-hydroxypropyl is also preferred. R³Compounds which are 5-quinolyl or 4-furazanyl are also preferred. More preferred compounds are those that combine the above characteristics. A particular isomer is also preferred.

The most preferred compound is 5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } -quinoline; in particular the (2R) -trans isomer thereof.

The compounds of the invention are chemosensitizers or synergists and may also be used as resistance modifiers. They can be used to treat multidrug resistance (i.e., after clinical resistance is apparent) and can also be used to enhance the activity of anticancer drugs by administration to the patient at the time of initiation of chemotherapy (i.e., prior to clinical resistance discovery). The compounds of the invention are also useful in the treatment of drug-resistant malaria.

The in vitro activity of chemosensitizers or potentiators, particularly the activity to treat multidrug resistance, is determined by MTT cell proliferation assays, such as the modified method described by Mosmann, T, "Rapid Colorimetric Assay For Cellular Growth and Surveillance," J.Immunol.meth., Vol.65,55-63 (1983). Another MTT Cell proliferation assay is described by Alley et al, "Feasibility of drug screening with circuits of Human Tumor Cell Lines Using Microcuulture Tetrazolium assay," Cancer Research, Vol.48,589-601 (1988).

The in vivo activity of a chemosensitizer or potentiator, particularly its activity in treating multidrug resistance, is determined as described by Slate and Michelson, "Drug resistance-and reverse variants: A company of Experimental dataWith Model precursors," J.Natl.cancer Inst., Vol.83,1574-1580 (1991). Other methods for in vivo testing are described by Sato et al, "circular-solution of multidug Resistance by a New Synthesized Quino-line deviation, MS-073," Cancer Research, Vol.51,2420-2424 (1991); tesuruo et al, "circulation of Vinceritine and mandrel Resistance in Vitro and in Vivo by Calcium Inf-lux Blockers," Cancer Research, Vol.43,2905-2910 (1983); and Tsouo et al, "topping of Vincristine Resistance in P388Leukmia in Vivo and in Vitro Enhanced cytoxicity of Vinristine and Vinblastine by Verapamide," Cancer Rese-arch, Vol.41,1967-1972 (1981).

The stability of such compounds in aqueous solution is determined by conventional methods, e.g., determining the amount of compound remaining in solution under various pH and temperature conditions.

The compounds of formula i are administered to a patient in a therapeutically effective dose, e.g., a dose sufficient to provide treatment for the disease state in question, particularly in combination with another active agent, preferably a cancer chemotherapeutic agent, particularly an agent selected from the foregoing, most preferably a cancer chemotherapeutic agent that produces significant clinical resistance in a mammal in need thereof. The compounds of the present invention and pharmaceutically acceptable salts thereof may be administered by any acceptable mode of administration having similar utility.

Although the dosage criteria for the compounds of the invention for use in humans have not been optimized, the daily dosage is generally from about 0.01 to 4.0mg per kg of body weight, preferably from about 0.1 to 2.0mg per kg of body weight, particularly preferably from about 0.3 to 1.0mg per kg of body weight. Thus, for a patient weighing 70kg, the amount of compound administered in the range of 0.7 to 280mg per day, preferably about 7.0 to 140mg per day, and particularly preferably about 21 to 70mg per day, depends on the patient and disease state, the extent of distress, the mode and time of administration (e.g., oral administration one day prior to cancer chemotherapy and intravenous administration during chemotherapy), and the judgment of the prescribing physician.

In the treatment of the above conditions using the compounds of the present invention, any pharmaceutically acceptable mode of administration may be used. The compounds of formula i may be administered alone or in combination with pharmaceutically acceptable excipients, including solid, semi-solid, liquid or aerosol forms, such as tablets, capsules, powders, liquid formulations, suspensions, suppositories, aerosols and the like. The compounds of formula i may also be administered in a sustained or controlled release form, including long acting injections, osmotic pumps, pills, transdermal patches (including electrical conductive) and the like, at a predetermined rate to prolong the administration time of the compound, preferably in a single administration of precise dosages used in unit dosage form. The compositions generally comprise conventional pharmaceutical carriers and excipients and the compounds of formula I and pharmaceutically acceptable salts thereof. In addition, other drugs, carriers, additives and the like, such as the above-mentioned cancer chemotherapeutic drugs and the like, may be additionally included in these compositions.

In general, pharmaceutically acceptable compositions will contain from about 0.1% to 90%, preferably from about 0.5% to 50%, by weight of the compound of formula I or salt thereof, with the remainder being suitable pharmaceutical excipients, carriers and the like, depending on the mode of administration to be effected.

For the above, a preferred mode of administration is oral, using conventional daily regimen, with appropriate adjustments depending on the degree of affliction. Such pharmaceutically acceptable non-toxic compositions for oral administration are prepared by the addition of any of the usual excipients, such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, Sodiumcrosscar mellose, glucose, gelatin, sucrose, magnesium carbonate, and the like. The composition comprises solution, suspension, tablet, dispersible tablet, pill, capsule, powder, sustained release preparation, etc.

The composition is preferably in the form of a pill or tablet. Thus, the composition contains, in addition to the active ingredient, diluents such as lactose, sucrose, dibasic calcium phosphate, and the like; lubricants such as magnesium stearate and the like; and binding agents such as starch, acacia, gelatin, polyvinylpyrrolidone, cellulose and derivatives thereof, and the like.

Liquid pharmaceutical compositions may be prepared, for example, by dissolving or dispersing the active compound as defined above and optional pharmaceutical additives in a carrier, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to form a solution or suspension. If desired, the pharmaceutical compositions may also contain minor amounts of non-toxic auxiliary substances such as lubricants, emulsifiers or co-solvents, pH buffers such as acetates, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and the like. The actual preparation of such dosage forms is known or simple to those of ordinary skill in the art; see, for example, Remington's pharmaceutical sciences, Mack Publishing Company, Easton, Pennsylvania,15th Edition, 1975. The composition or the prescription to be administered should in any case contain an amount of active compound sufficient to alleviate the symptoms of the patient to be treated.

The active ingredient accounts for 0.005-95%, and other dosage forms or compositions composed of nontoxic carriers can also be prepared.

Pharmaceutically acceptable non-toxic compositions for oral administration may be prepared by the addition of conventional excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, talcum, cellulose derivatives, Sodium crosscar mellose, glucose, sucrose, magnesium carbonate, Sodium saccharin and the like. These compositions may take the form of solutions, suspensions, tablets, capsules, powders, depot formulations, and the like. The composition contains 0.01% -95% of active ingredient, preferably 0.1-50%.

For solid dosage forms, solutions or suspensions in propylene or ethylene carbonate and mixtures thereof, and vegetable oils or triglycerides, gelatin capsules are preferred. The solution and the preparation and encapsulation thereof are already described in U.S.4,328, 245; 4,409,239 and 4,410,545. For liquid dosage forms, for example, a solution of polyethylene glycol may be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier, such as water, to facilitate administration.

Alternatively, liquid or semisolid oral formulations can also be prepared by dissolving or dispersing the active compound or and salt in vegetable oils, glycols, triglycerides, propylene glycerides (e.g., propylene carbonate), ethylene carbonate, and mixtures thereof, and then filling the solution or suspension into hard or soft capsules.

Other useful formulations also include those disclosed in U.S.281819 and 4,358,603.

Suitable for oral and parenteral administration is a further advantage of the present invention, since the compounds of formula I are much more stable than the MS-073 compounds.

Parenteral administration is usually by injection or subcutaneous, transdermal or intravenous administration. Injectables can be prepared by conventional methods, such as liquid solutions or suspensions, solid forms suitable for constitution with a solution or suspension prior to injection, or injectables in the form of emulsions. Suitable excipients are water, saline, dextrose, glycerol, ethanol, and the like. In addition, if desired, the pharmaceutical compositions may also contain minor amounts of non-toxic additives such as wetting or emulsifying agents, pH buffering agents, solubilizing agents, and others such as sodium acetate, sodium sorbitan monolaurate, triethanolamine oleate, cyclodextrins, and the like.

A more recent approach to parenteral administration is to implant sustained or long acting systems that maintain the dose at a constant level, see U.S.3,710,795.

In compositions for parenteral administration, the percentage of active substance depends on its specific properties and the activity of the compound and the needs of the patient. In solution, percentages of active ingredient between 0.01 and 10% are possible, but if the composition is in solid form, the percentages will be slightly higher, since dilution to the above percentages is required in preparation. Preferably, the composition contains 0.2-2% of active substance in solution.

Nasal drops of the nasal solutions containing the active compounds may also be administered, alone or in combination with other pharmaceutically acceptable excipients.

The formulations of the active compounds or their salts may also be administered alone or in combination with an inert carrier such as lactose to the respiratory tract, for example, as an aerosol or spray, or as a fine powder for nasal inhalation. In this case, the particles of the formulation should have a diameter of less than 50 microns, preferably less than 10 microns.

The following preparations and examples are given to enable one of ordinary skill in the art to more clearly understand and practice the present invention, and are intended to be illustrative and representative of the present invention, and should not be construed as limiting the scope of the present invention. Example 110, 11-Difluoromethylene dibenzosuberone 1 A.R¹And R²A compound of formula 1 which is F

A solution of 350g of sodium chlorodifluoroacetate in 1400ml of diglyme was added dropwise over 6 hours, with stirring and under a nitrogen atmosphere, to a solution of dibenzosuberone (25g) in diglyme (500ml), the reaction temperature being maintained at 160 ℃ and 165 ℃, and the cooled reaction mixture was poured into water (1)8 l), extraction with 1.8 l of diethyl ether, washing of the organic phase with water and Na₂SO₄Drying and subsequent evaporation, the residue is recrystallized from ethanol and then acetone/hexane to give 14g of 10, 11-difluoromethylenedibenzosuberone, mp149.6 ℃. The combined mother liquors were flash chromatographed on silica gel column, eluting with 20% acetone/hexane to give 6.5g additional desired product. 1B. variation R¹And R²A compound of formula 1

According to the preparation method of A, the sodium chlorodifluoroacetate is replaced by the following substances: a. methyl trichloroacetate, b. methyl tribromoacetate, and c. sodium dichlorofluoroacetate;

the following compounds were obtained: 10, 11-dichloromethylene dibenzosuberone b.10, 11-dibromomethylene dibenzosuberone and c.10, 11-chlorofluoromethylene dibenzosuberone; example 210, 11-Difluoromethylene dibenzosuberol 2 A.R¹And R²A compound of formula 3 which is fluorine

A solution of 10, 11-difluoromethylenedibenzosuberone (20.4g) in 900ml THF/MeOH (1: 2) was cooled in an ice bath. 12g of sodium borohydride were added in portions, the cold bath was removed and the reaction mixture was stirred at room temperature for 2 hours and poured into water. The product is filtered off, washed with water and then dried, giving 20g of 10, 11-difluoromethylenedibenzosuberol, mp230.1-230.6 ℃. 2B. variation R¹And R²A compound of formula 2

The preparation according to part a was carried out using the following materials in place of 10, 11-difluoromethylenedibenzosuberone: 10, 11-dichloromethylene dibenzosuberone, b.10, 11-dibromomethylene dibenzosuberone, c.10, 11-methylene dibenzosuberone, and d.10, 11-chlorofluoromethylene dibenzosuberone;

the following compounds can be obtained separately: 10, 11-dichloromethylene dibenzosubenol, b.10, 11-dibromomethylene dibenzosubenol, c.10, 11-methylene dibenzosubenol, and d.10, 11-chlorofluoro-methylene dibenzosubenol. Example 3 cis and trans-1- (10, 11-Difluoromethylene dibenzosuberyl-5-yl) -4-formylpiperazine 3 A.R¹And R²Compounds of formula 3 wherein R is fluoro and R is formyl

To a solution of 10, 11-difluoromethylenedibenzosuberyl alcohol (5.2g) in 70ml of dioxane, which had been cooled in an ice bath in advance, thionyl chloride (4.5ml) was added dropwise. The reaction temperature was raised to 50 ℃. And held for 4 hours. The reaction mixture was evaporated to dryness to give a mixture of cis and trans-5-chloro-10, 11-difluoromethylenedibenzosuberane (5.7g), which was dissolved in acetonitrile (200ml) and 10ml of 1-piperazinecarboxaldehyde were added and the mixture was dried over N₂Stirring under gas at 100 deg.C (water bath temperature) for 20 hr, then evaporating to dryness, and separating the residue with NaHCO₃Partitioning the aqueous solution with ethyl acetate, washing the organic phase with water, K₂CO₃Drying, evaporation to dryness and flash chromatography of the residue on silica gel (30% acetone/hexanes) afforded cis-1-10, 11-difluoromethylenedibenzocyclohept-5-yl) -4-formylpiperazine (2.4g), mp213 ℃, and trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) -4-formylpiperazine (2.6g), mp238 ℃. 3B. variation R¹And R²A compound of formula 3

According to the preparation method of A,10, 11-difluoromethylene dibenzosuberol is replaced by the following substances: 10, 11-dichloromethylene dibenzosubenol, b.10, 11-dibromomethylene dibenzosubenol, c.10, 11-methylene dibenzosubenol, and d.10, 11-chlorofluoro-methylene dibenzosubenol;

the following compounds were obtained: a1. trans-1- (10, 11-dichloromethylene dibenzosuberyl-5-yl) -4-formylpiperazine,

mp205 ℃ and a 2. cis-1- (10, 11-dichloromethylenedibenzosuben-5-yl) -4-formylpiperazine, b 1. trans-1- (10, 11-dibromomethylenedibenzosuben-5-yl) -4-formylpiperazine, b2. cis-1- (10, 11-dibromomethylenedibenzosuben-5-yl) -4-formylpiperazine, c 1. trans-1- (10, 11-methylenedibenzosuben-5-yl) -4-formylpiperazine, mp

195 deg.C, c2. cis-1- (10, 11-methylenedibenzosuberyl-5-yl) -4-formylpiperazine, d1. trans-1- (10, 11-chlorofluoromethylenedibenzosuberyl-5-yl) -4-formylpiperazine

Oxazines, and d 2. cis-1- (10, 11-chlorofluoromethylenedibenzosuberyl-5-yl) -4-formylpiperazine

And (3) an oxazine. Example 4 Trans-1- (10, 11-Difluoromethylenedibenzocyclohept-5-yl) piperazine 4 A.R¹And R²A compound of formula 4 which is fluorine

A solution of 2.55g of trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) -4-formylpiperazine and 100ml of potassium hydroxide (3.0g) in ethanol/water (9: 1) was refluxed for 1 hour and then cooled. The cooled reaction mixture was concentrated, diluted with water, extracted with ethyl acetate, K₂CO₃Drying and evaporation of the dried organic phase gave trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine (2.35g), mp131 ℃. 4B. variation R¹And R²A compound of formula 4

According to the preparation of A, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) -4-formylpiperazine was replaced by: a. cis-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) -4-formylpiperazine, b, trans-1- (10, 11-dichloromethylenedibenzosuberyl-5-yl) -4-formylpiperazine, c, cis-1- (10, 11-dichloromethylenedibenzosuberyl-5-yl) -4-formylpiperazine, d, trans-1- (10, 11-dibromomethylenedibenzosuberyl-5-yl) -4-formylpiperazine, e, cis-1- (10, 11-dibromomethylenedibenzosuberyl-5-yl) -4-formylpiperazine, f, trans-1- (10, 11-methylenedibenzosuberyl-5-yl) -4-formylpiperazine, g.cis-1- (10, 11-methylenedibenzosuberyl-5-yl) -4-formylpiperazine, h.trans-1- (10, 11-chlorofluoromethylenedibenzosuberyl-5-yl) -4-formylpiperazine,

cis-1- (10, 11-chlorofluoromethylenedibenzosuberyl-5-yl) -4-formylpiperazine;

the following compounds can be obtained separately: a. cis-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine, mp

225.5 ℃, b.trans-1- (10, 11-dichloromethylenedibenzosuberyl-5-yl) piperazine, mp199 ℃, c, cis-1- (10, 11-dichloromethylenedibenzosube-5-yl) piperazine, d, trans-1- (10, 11-dibromomethylenedibenzosube-5-yl) piperazine, e, cis-1- (10, 11-dibromomethylenedibenzosube-sube-5-yl) piperazine, f, trans-1- (10, 11-methylenedibenzosube-5-yl) piperazine, mp103 ℃, g, cis-1- (10, 11-methylenedibenzosube-5-yl) piperazine, h, trans-1- (10, 11-chlorofluoromethylenedibenzosube-5-yl) piperazine, cis-1- (10, 11-chlorofluoromethylenedibenzocyclohept-5-yl) piperazine, example 51- (4-benzofurazanyloxy) -2, 3-epoxypropane 5 A.R³Compounds of formula 8 which are benzofurazanyl and n is 1

620mg of sodium hydride (60% oil dispersion) was added portionwise to a solution of 1.74g of benzofurazan-4-ol in dimethylformamide (30ml), the mixture was heated at 50 ℃ for 30 minutes, 3-bromo-1, 2-epoxypropane (1.6ml) was added, and the mixture was heated at 60 ℃ for 2 hours. The reaction mixture was poured into water, extracted with ethyl acetate, the organic phase washed with water, Na₂SO₄Dried and evaporated. Chromatography of the residue on silica gel (50% ethyl acetate in hexanes) afforded 1- (4-benzofurazanyloxy) -2, 3-epoxypropane (1.6g), mp75 ℃. 5B. variation R³And n are compounds of formula 8

According to the preparation method of part A, benzofurazan-4-ol and 3-bromo-1, 2-epoxypropane are replaced by the following substances: a. quinoline-5-ol and 1-chloro-3, 4-butylene oxide, b.2-nitrophenol and 3-bromo-1, 2-propylene oxide, c.2-chlorophenol and 3-bromo-1, 2-propylene oxide, d.2-difluoromethoxyphenol and 3-bromo-1, 2-propylene oxide, e.g. pyridin-3-ol and 3-bromo-1, 2-propylene oxide, and f.quinoline-5-ol and 3-bromo-1, 2-propylene oxide;

the following compounds can be obtained separately: 1- (5-quinolinyloxy) -3, 4-epoxybutane, b.1- (2-nitrophenoxy) -2, 3-epoxypropane, c.1- (2-chlorophenoxy) -2, 3-epoxypropane, d.1- (2-difluoromethoxyphenoxy) -2, 3-epoxypropane, e.1- (3-pyridinyloxy) -2, 3-epoxypropane, and f.1- (5-quinolinyloxy) -2, 3-epoxypropane. 5℃ variation R³And n are compounds of formula 9

According to the preparation method of part A, benzofurazan-4-ol and 3-bromo-1, 2-epoxypropane are replaced by the following substances: a. quinolin-5-ol and 1-bromo-3-chloropropane, b. quinolin-5-ol and 1-bromo-4-chlorobutane, c.2-nitrophenol and 1-bromo-3-chloropropane;

the following compounds can be obtained separately: 1- (5-quinolinyloxy) -3-chloropropane, b.1- (5-quinolinyloxy) -4-chlorobutane, and c.1- (2-nitrophenoxy) -3-chloropropane. Example 6(2R, S) -trans-5- {3- [4- (10, 11-Difluoromethylene dibenzocyclohept-5-yl) piperazin-1-yl]-2-hydroxypropoxy } quinoline 6 A.R¹And R²Is fluorine, A is (2R, S) -hydroxypropyl, and R is³Compounds of formula I which are 5-quinolyl

A solution of 586mg of 1- (5-quinolinyloxy) -2, 3-epoxypropane and trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine (950mg) in 20ml of isopropanol was refluxed for 20 hours, the reaction mixture was evaporated to dryness, and the residue was chromatographed on silica gel (70: 30: 1 ethyl acetate/hexane/triethylamine) to give (2R, S) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl]-2-hydroxypropoxy } quinoline (1.33g) which was reacted with 3mol equiv of HCl and converted to the trihydrochloride salt, mp193.5 ℃. 6B. variation R¹、R²、R³And A is an isomer of formula I

According to the preparation method of part A, 1- (5-quinolinyloxy) -2, 3-epoxypropane of trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine-was replaced with the following: a. cis-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (5-quinoline

Oxy) -2, 3-epoxypropane, b, trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and (2R) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, c.trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and (2S) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, d, cis-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and (2R) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, e.cis-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and (2S) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, f, trans-1- (10, 11-dichloromethylene dibenzosuberyl-5-yl) piperazine and 1- (5-quinoline

Oxy) -2, 3-epoxypropane, g, trans-1- (10, 11-dichloromethylene dibenzosuberyl-5-yl) piperazine and (2R) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, h.trans-1- (10, 11-dichloromethylenedibenzosuberyl-5-yl) piperazine and (2S) -1- (5)

-quinolinyloxy) -2, 3-epoxypropane, i.trans-1- (10, 11-methylenedibenzosuben-5-yl) piperazine and 1- (5-quinolinyloxy)

-2, 3-epoxypropane, j, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (4-benzo

Furazanyloxy) -2, 3-epoxypropane, k, cis-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and 1- (4-benzo

Furazanyloxy) -2, 3-epoxypropane, l, trans-1- (10, 11-dichloromethylene dibenzosuberyl-5-yl) piperazine and 1- (4-benzo

Furazanyloxy) -2, 3-epoxypropane, m, trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and 1- (2-nitro-2

Phenoxy) -2, 3-epoxypropane, n, trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and 1- (2-chlorobenzene

Oxy) -2, 3-epoxypropane, o, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (2-difluoro

Methoxyphenoxy) 2, 3-epoxypropane, p, trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and 1- (3-pyridine

Oxy) -2, 3-epoxypropane, q, trans-1- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine and 1- (5-quinoline

Oxy) -3-chloropropane, r, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (5-quinoline

Oxy) -4-chlorobutane, s, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (2-nitro-n-ylbenzocyclohepta-5-yl)

Phenoxy) -3-chloropropane, and t, trans-1- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine and 1- (5-quinoline

Oxy) -3, 4-epoxybutane,

the following compounds can be obtained separately: (2R, S) -cis-5- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperacillin

Oxazin-1-yl ] -2-hydroxypropoxy } quinoline, mp208 ℃ (trihydrochloride), b. (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp190 ℃ (trihydrochloride), c. (2S) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp195 ℃ (trihydrochloride), d. (2R) -cis-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp193 ℃ (trihydrochloride), e. (2S) -cis-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp188.5 ℃ (trihydrochloride), f. (2R, S) -trans-5- {3- [4- (10, 11-dichloromethylene dibenzocyclohept-5-yl) piper-ine

Oxazin-1-yl ] -2-hydroxypropoxy } quinoline, mp195 ℃ (trihydrochloride), g. (2R) -trans-5- {3- [4- (10, 11-dichloromethylene dibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp218 ℃ (trihydrochloride), h. (2S) -trans-5- {3- [4- (10, 11-dichloromethylenedibenzocyclohept-5-yl) piperazine

-1-yl ] -2-hydroxypropoxy } quinoline, mp215 ℃ (trihydrochloride salt), i. (2R, S) -trans-5- {3- [4- (10, 11-methylenedibenzosuberyl-5-yl) piperazine-1

-yl ] -2-hydroxypropoxy } quinoline, melting range: 874-99.3 deg.C; MS: molecular ion

Peak =491, j. (2R, S) -trans-4- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piper-ine

Oxazin-1-yl ] -2-hydroxypropoxy } benzofurazan, mp186 ℃ (dihydrochloride), k. (2R, S) -cis-4- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazine

Oxazin-1-yl ] -2-hydroxypropoxy } benzofurazan, mp188 ℃ (dihydrochloride), l. (2R, S) -trans-4- {3- [4- (10, 11-dichloromethylenedibenzosuberyl-5-yl) piperazine

Oxazin-1-yl ] -2-hydroxypropoxy } benzofurazan, m. (2R, S) -trans-1- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine

Oxazin-1-yl ] -2-hydroxypropoxy } -2-nitrobenzene, n. (2R, S) -trans-1- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piper-ine

Oxazin-1-yl ] -2-hydroxypropoxy } -2-chlorobenzene, o. (2R, S) -trans-1- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piper-ine

Oxazin-1-yl ] -2-hydroxypropoxy } -2-difluoromethoxybenzene, p. (2R, S) -trans-3- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piper-ine

Oxazin-1-yl ] -2-hydroxypropoxy } pyridine, q. (2R, S) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piperazine

Oxazin-1-yl-propoxy-quinoline, R. (2R, S) -trans-5- {4- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piper-ine

Oxazin-1-yl ] butoxy } quinoline, S. (2R, S) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piper-ine

Oxazin-1-yl ] propoxy } -2-nitrobenzene, and t. (2R, S) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzosuberyl-5-yl) piper-ine

Oxazin-1-yl ] -3-hydroxybutoxy } quinoline. Example 7

This example describes the preparation of a representative pharmaceutical formulation for oral administration containing a compound of formula I, such as (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline.

Amount of ingredient in each tablet (mg)

Active compound 20

Spray dried lactose 148

Magnesium stearate 2

Mixing the above materials, and encapsulating. If the active substance trihydrochloride salt is used, 243mg of salt are used.

Other compounds of formula I, such as those prepared according to the methods described in examples 1-6, may also be used as active substances in the preparation of formulations for oral administration in this example. Example 8

This example describes another representative method for preparing a pharmaceutical formulation for oral administration containing a compound of formula I, such as (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline.

Amount of ingredient in each tablet (mg)

Active compound 400

Corn starch 50

Lactose 145

Magnesium stearate 5

The above ingredients were mixed thoroughly and compressed into a scored tablet, which, if its trihydrochloride salt was used, required 486 mg.

Other compounds of formula I, such as those prepared according to the methods described in examples 1-6, may also be used as active substances in the preparation of oral formulations in this example. Example 9

This example describes the preparation of a representative pharmaceutical formulation containing an active compound of formula I, such as (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline.

A suspension for oral administration comprising the following components was prepared as follows:

amount of ingredients

Active substance 1.0g

Fumaric acid 0.5g

Sodium chloride 2.0g

methyl paraben 0.1g

Granulated sugar 25.5g

Sorbitol (70% solution) 12.85g

Veegum K(Vanderbilt Co.) 1.0g

0.035ml flavoring agent

Colorant 0.5mg

Distilled water of proper amount to 100ml

If its trihydrochloride salt is used to prepare a suspension, 1.215g of salt is required. Other compounds of formula I, such as those prepared according to the methods of examples 1-6, may also be used as the active compound in the oral administration formulations of this example. Example 10

This example describes the preparation of a representative pharmaceutical formulation containing a compound of formula I, such as (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline.

A method for preparing injection with pH of 4 comprises the following components:

the amount of the components

Active Compound 0.2g

Sodium acetate buffer (0.4M) 2.0ml

HCl (1N) in appropriate amount to pH4

Appropriate amount of water (distilled water, sterile) to 20ml

If the trihydrochloride salt of the active compound is used to prepare the injection, 243g of salt are used. Other compounds of formula I, such as those prepared according to the methods of examples 1-6, may also be used as active compounds in the preparation of the injectables of this example. Example 11

This example describes the preparation of a representative pharmaceutical formulation containing an active compound of formula I, such as (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline.

A suppository weighing 2.5g is prepared containing the following components:

active Compound 500mg

witepsol H-15^*Equilibrium (triglycerides of saturated vegetable fatty acids, Riches-Nelson, inc., New York, n.y.)

If the trihydrochloride salt of the active substance is used, 607mg of the salt are used. Other compounds of formula I, prepared according to the methods of examples 1-6, may also be used as active substances in the preparation of the suppositories of this example. EXAMPLE 12 determination of stability under acidic pH conditions

The test compound (15. mu.g) was dissolved in 3ml of 0.01N HCl (pH2) and incubated at 37 ℃. At various times, 10. mu.l of each sample was withdrawn and injected into a 3 μm Pecosphere C-18 column (3.3X 0.48cm) for HPLC analysis (mobile phase: 35% acetonitrile/18% tetrahydrofuran/47% potassium dihydrogen phosphate, mobile phase containing 4mM N, N-dimethyloctylamine; flow rate 1.0 ml/min). The test compounds and their degradation products were detected at 240nm of UV absorption. The disappearance of the parent compound is expressed as a percent peak height of zero with respect to time, from which the value of t1/2 can be determined.

MS-073(US5,112,817), t1/2=15 minutes,

(2R, S) -trans-5- {3- [4- (10, 11-methylenedibenzosuberyl-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline, t1/2=2.5 hours,

(2R, S) -5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline, t1/2= > 72 hours, and

(2R, S) -trans-5- {3- [4- (10, 11-chloromethylene dibenzosube-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline, t1/2= > 72 hours.

The compounds of the present invention exhibit significantly better stability than MS-703 at acidic pH as determined by the method. Example 13 in vitro Activity assay Using the MTT method

The method is a modification of the test method described by Mosmann, T.in "Rapid Colorimetric Assay for cellular Growth And solar review: Application to promotion And cytoxcity assays," J.Immunol.meth., Vol.65,55-63 (1983).

Will contain multidrug resistant CH^RC5 hamster cell (about 2X 10)⁵Cell number/ml) was combined with the medium containing the test compound or vehicle control to form a suspension in the presence or absence of adriamycin ((1. mu.g/ml). 0.1ml portions of the cell suspension were taken and placed in 8 wells of each of three 96-well microtiter plates. The culture was carried out in a tissue culture incubator at 37 ℃ and one plate was taken out at each of the following different times: 24 hours, 48 hours and 72 hours. After removal of the incubator, 3- [4, 5-dimethylthiazol-2-yl ] was added to each well of the plate]2, 5-Diphenylbromotetrazole, MTT (10. mu.l, from 5mg/ml of a pre-prepared solution in phosphate buffered saline), and then returned to the incubator for 3 hours.

Formazan crystals produced by mitochondrial enzyme activity in living cells were solubilized by removing the medium and adding DMSO (15. mu.l/well) and mixing well on a shaker. A570 (reference wavelength 650nm) was read on a Molecular Devices microdisk reader and the results were expressed as a percentage of daily steroid control or Adrias control or plotted against A570 control time.

When tested by this method, the compounds of the present invention exhibit activity. In particular, the compound (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } -quinoline is more than 3 times more active than MS-73; the corresponding (2S) -trans isomer was 1.7 times as active as MS-73, while the (2R, S) -trans mixture was 1.6 times as active as MS-73. EXAMPLE 14 measurement of in vivo Activity Using MDR assay

The present method is an improvement of the tests described by Slate and Michelson, see "drug resistance reverse Strategies: A company of Experiment-al Data With models Predictions," J.Natl.cancer Inst., Vol83,1574-1580 (1991).

Mice (B6D2F1, female, 7-8 weeks, ca. 20g, Jackson laboratory-r)y) weighed and randomly divided into 6-7 pieces per group. On day 0, mice were injected intraperitoneally with o.2mlP388/ADR multidrug-resistant murine leukocytes, 2.4X 10⁷cells/ml. Two hours later, each mouse was implanted peritoneally with an Alzet7 day micropump (Model 2001, Alza Corporation, Palo Alto, CA) containing: control (DMSO/PBS) plus doxorubicin (3mg/kg/day), test compound alone (30mg/kg/day) or doxorubicin (3mg/kg/day) plus test compound (0.3, 3, 10 and 30 mg/kg/day). Mice were monitored daily and deaths were recorded starting on day 7. An increase in survival time compared to the control and adriamycin groups indicates activity.

When tested by this method, the compounds of the invention show activity. In particular, the compound (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline is 4 times more active than MS-73; the corresponding (2S) -trans isomer was 1.3 times more active than MS-73.

Also, the compound (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline is 3-fold more active than MS-73 in the human uterine sarcoma assay. EXAMPLE 15 toxicity of Compounds of formula I

Mice received (2R) -trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropoxy } quinoline by intravenous pharmaceutical injection once daily for 2 weeks at a dose of 0 (control: 5% w/v mannitol in purified water), 10, 30, and 100 mg/kg. All mice survived in the 0-, 10-and 30-mg/kg groups. All mice given 100mg/kg showed clonic convulsions after the first dose and died. The cause of death is pending.

Claims (9)

1. A combination product for simultaneous, separate or subsequent use in the treatment of cancer, wherein the product comprises a compound of the formula or a pharmaceutically acceptable salt thereof and a cancer chemotherapeutic agent

Wherein:

a is-CH₂-CH₂-,-CH₂-CHR^a-CH₂-, wherein R^aIs H, OH or a lower acyloxy group of the formula-O-C (= O) -R ', wherein R' is cyclic, branched or straight C_1-6One is monovalentAlkyl, or-CH₂-CHR^a-CHR^b-CH₂-, wherein R^aOr R^bOne is H, OH, or a lower acyloxy group of the formula-O-C (= O) -R ', where R' is cyclic, branched or straight C_1-6A monovalent alkyl group, the other being hydrogen;

R¹is H, F, Cl or Br;

R²is H, F, Cl or Br; and

R³is heteroaryl having 2 condensed rings and 1-3 heteroatoms, wherein said heteroatoms are N, O or S.

2. A combination product according to claim 1 wherein R is³Is quinolinyl or benzofurazanyl.

3. A complexing product according to claim 2 wherein R³Is a 5-quinolyl group.

4. A complexing product according to either one of claims 1 or 3 wherein R^aOr R^bIs OH.

5. A combination product according to claim 4 wherein A is-CH₂-CHR^a-CH₂-and R^aIs OH.

6. The combination product of claim 1 wherein said compound, or pharmaceutically acceptable salt thereof, has the formula

7. The combination product of claim 1 wherein said compound, or pharmaceutically acceptable salt thereof, has the formula

8. The complexing agent product of claim 1 wherein said complexing agent product,wherein said compound or pharmaceutically acceptable salt thereof has the following structural formula

9. The combination as defined in claim 6, wherein the compound or its salt is (2S-trans-5- {3- [4- (10, 11-difluoromethylenedibenzocyclohept-5-yl) piperazin-1-yl ] -2-hydroxypropyl } quinoline or a pharmaceutically acceptable salt thereof.