WO2008124145A1 - Composés stéroïdes, compositions et procédés de traitement - Google Patents

Composés stéroïdes, compositions et procédés de traitement Download PDF

Info

Publication number
WO2008124145A1
WO2008124145A1 PCT/US2008/004541 US2008004541W WO2008124145A1 WO 2008124145 A1 WO2008124145 A1 WO 2008124145A1 US 2008004541 W US2008004541 W US 2008004541W WO 2008124145 A1 WO2008124145 A1 WO 2008124145A1
Authority
WO
WIPO (PCT)
Prior art keywords
independently
alkyl
compound
aryl
cycloalkyl
Prior art date
Application number
PCT/US2008/004541
Other languages
English (en)
Inventor
Yves Pommier
Christophe Marchand
Smitha Antony
Thomas Dexheimer
Original Assignee
Government Of The U.S.A., As Represented By The Secretary, Department Of Health And Human Services
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Government Of The U.S.A., As Represented By The Secretary, Department Of Health And Human Services filed Critical Government Of The U.S.A., As Represented By The Secretary, Department Of Health And Human Services
Publication of WO2008124145A1 publication Critical patent/WO2008124145A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Cancer in all its manifestations, remains a devastating disorder. Although cancer is commonly considered to be a single disease, it actually comprises a family of diseases wherein normal cell differentiation is modified so that it becomes abnormal and uncontrolled. As a result, these malignant cells rapidly proliferate. Eventually, the cells spread or metastasize from their origin and colonize other organs, eventually killing their host. Due to the wide variety of cancers presently observed, numerous strategies have been developed to destroy cancer within the body. Cancer is generally characterized by the presence of endogenous DNA damage. However, while DNA damage underlies carcinogenesis, it has also been exploited as a means to treat cancer. For example, the cytotoxic effects of the two main therapeutic modalities that are clinically used to treat malignances (i.e.
  • DNA damage is also responsible for many of the side effects of these therapies, such as bone marrow suppression, gastrointestinal toxicities, and hair loss.
  • the sensitivity of cancer cells to DNA damaging agents is probably related to intrinsic deficiencies in DNA repair and checkpoint mechanisms.
  • the capacity of cancer cells to recognize DNA damage and initiate DNA repair is a key mechanism for therapeutic resistance to chemotherapy. Therefore, the targeting of DNA repair enzymes for anticancer therapeutic intervention can be used as a strategy to potentiate the cytotoxicity of the currently available DNA damaging agents toward cancer cells.
  • cancer is treated by chemotherapy, in which highly toxic chemicals are given to the patient, or by radiotherapy, in which toxic doses of radiation are directed at the patient.
  • chemotherapy in which highly toxic chemicals are given to the patient
  • radiotherapy in which toxic doses of radiation are directed at the patient.
  • these "cytotoxic" treatments also kill extraordinary numbers of healthy cells, causing the patient to experience acute debilitating symptoms including nausea, diarrhea, hypersensitivity to light, hair loss, etc.
  • the side effects of these cytotoxic compounds limits the frequency and dosage at which they can be administered.
  • Such disabling side effects can be mitigated to some degree by using compounds that selectively target cycling cells, i.e., interfering with DNA replication or other growth processes in cells that are actively reproducing. Since cancer cells are characterized by their extraordinary ability to proliferate, such protocols preferentially kill a larger proportion of cancer cells in comparison to healthy cells, but cytotoxicity and ancillary sickness remains a problem.
  • Another strategy for controlling cancer involves the use of signal transduction pathways in malignant cells to "turn off' their uncontrolled proliferation, or alternatively, instruct such cells to undergo apoptosis.
  • Such methods of treating cancer are promising but a substantial amount of research is needed in order to make these methods viable alternatives.
  • Cancer has been typically treated with surgery, radiation and chemotherapy, alone or in conjunction with various therapies employing drugs, biologic agents, antibodies, and radioactive immunoconjugates, among others.
  • the common goal of cancer treatment has been, and continues to be, the elimination or amelioration of cancerous tumors and cells with minimal unpleasant or life-threatening side effects, due to toxicity to normal tissues and cells.
  • these goals remain largely unmet.
  • TDPs tyrosyl-DNA phosphodiesterases
  • TDPl tyrosyl-DNA phosphodiesterase 1
  • the invention provides a method of inhibiting Tdpl activity in a subject, the method comprising administering to the subject a compound of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(R b )N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -0R b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R 3 , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted; wherein each W is independently C, N, CR a , or NR b ; each R
  • the invention provides a method of treating a Tdpl -related disorder in a subject, comprising administering to said subject in need thereof, an effective amount of a compound of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R 3 , -NR b SO 2 R 3 , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted; wherein each W is independently C, N, CR a , or NR b ; each R a is
  • the invention provides a method of treating cancer in a subject identified as in need of such treatment, the method comprising administering to said subject an effective amount of a compound of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R 3 , -OS(O) 2 R 3 , -OP(O)R 2 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted; wherein each W is independently C, N, CR a , or NR b ; each R a
  • the invention provides for the use of a steroid compound in the manufacture of a medicament for inhibiting or reducing cancer in a patient, the compound being of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is 0-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b S0 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted; wherein each W is independently C, N, CR a , or NR b ; each R
  • the invention provides a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkyl carbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R 3 , -NR b SO 2 R 3 , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR a , or NR b ; each R a is independently, H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, haloalkyl, -OR b , -SR b , -NR b R b , hydroxylalkyl, alkylcarbonylalkyl, mercaptoalkyl, aminoalkyl, sulfonylalkyl, sulfonylaryl, or thioalkoxy; each of which may be optionally substituted; each R b is independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, or heteroaryl; each of which may be
  • the invention provides for a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR 3 , or NR b
  • the invention provides for a kit comprising an effective amount of a steroid compound in unit dosage form, together with instructions for administering the steroid compound to a subject suffering from cancer, wherein the compound is a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(0)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R a R a , each of which may be optionally substituted;
  • a particularly preferred compound of the invention has the following structure A:
  • Compound A (also referred to herein as NSC 88915) is an effective inhibitor of Tdpl .
  • Compound A can be employed in therapeutic methods disclosed herein, including to treat cancer.
  • Compound A can inhibit the activity of recombinant human Tdpl at low micromolar concentrations both with single and double-stranded DNA substrates. Additionally, based on our analysis including surface plasmon resonance analysis and molecular modeling, it is proposed without limitation that Compound A (NSC 88915) is a competitive Tdpl inhibitor, which can mimic the binding mode of the Tdpl substrate.
  • compounds disclosed herein have use against cancers (e.g. solid tumors) with preexisting DNA repair and cell cycle checkpoint deficiencies.
  • FIG. 1 A) Schematic representation of the human Tdpl domain structure. The N- terminal and C-terminal domains correspond to residues 1-350 and 351-608, respectively. Positions of the "HKN” motifs and NLS (nuclear localization signals) motifs are shown. Arrows indicate active site residues. B) Ribbon diagram of Tdpl structure ( ⁇ 148). C) Structure of the Tdpl-vanadate-peptide-DNA complex (INOP). Tdpl is shown as a molecular surface and the vanadate-peptide-DNA substrate mimic is shown as ball-and-stick structures with the DNA, the vanadate, and the Topi -derived peptide shown.
  • FIG. 1 Proposed reaction mechanism for human Tdpl .
  • A) The first step of the reaction involves the nucleophilic attack of the phosphodiester-containing substrate by the imidazole N ⁇ 2 atom of His263. H493 donates a proton to the tyrosyl moiety of the leaving group. The phosphohistidine intermediate is shown in (B). Interactions between the non-bridging oxygens and the active site lysine residues (Lys265 and Lys495) serve to stabilize the transition intermediate.
  • C) The next step of the reaction involves a nucleophilic attack by a water molecule activated by H493, generating final 3 '-phosphate product and free Tdpl (D).
  • FIG. 3 Outline of the substrate binding pocket and the different substrates processed by Tdpl .
  • A) Molecular surface of Tdpl .
  • the black dotted lines define the substrate binding pocket.
  • the R- group refers to the structures shown in (C) and (D). The arrow indicates the Tdpl cleavage site.
  • C) and D) illustrate the physiological and non-physiological substrates of Tdpl.
  • E) and F) show both favorable and unfavorable the Tdpl substrates.
  • the black circle represents a 3 '-end phosphotyrosine.
  • E) (i) long single-strand, (ii) tailed duplex, (iii) gapped duplex.
  • F) (i) short single-strand, (ii) nicked duplex, (iii) full-length Topi .
  • FIG. 4 Redundant pathways involved the repair of Topi cleavage complexes in mammalian cells.
  • Figure 5 Principles of the BVTM Technology. Upper panel Structure of a ruthenium- containing BV-T AGTM label designed to be incorporated into oligonucleotides. Lower panel/Diagram of the excitation/emission mechanism of the electrochemiluminescent BV- TAGTM.
  • FIG. 6 Coupling reaction using a BV-TAGTM NHS ester and a Tdpl DNA substrate bearing a phosphotyrosine group at its 3' end. After coupling, the BV-Tag is attached to the phosphotyrosine and a succinimide getup is released. The labeled material is then purified on an Oligo Spin Column (BioRad, Hercules, CA).
  • Tdpl catalytic reaction leading to the process of the Tdpl -BV-Tag DNA substrate.
  • Tdpl cleaves the phosphotyrosine removing the tyrosine-BV-Tag group and leaving a 3' phosphate on the DNA. This leads to a loss of the electrochemiluminescence signal. Positive hits for potential Tdpl inhibitors prevent this loss of signal.
  • FIG. 9 Summary graph representing the screening results of the 1981 -compound DTP Diversity set tested at 10 ⁇ M. Each dot indicates a signal value for a tested sample.
  • the dashes line represent 50% inhibition.
  • NSC 88915 gives a signal value of 16801 (indicated by an arrow) which corresponds to 100% inhibition at 10 ⁇ M.
  • FIG. 10 Steroid derivatives confirmed in a gel -based assay as Tdpl inhibitors. Two of them (NSC 88915 and 109131) were counter-screened against tile enzyme Apel and were shown to be selective inhibitors of Tdpl .
  • t Figure 11 A. Representative gel showing inhibition of Tdpl activity by Compound A (NSC 88915) on three different substrates (i.e. single stranded, blunt duplex and 5 '-tailed duplex containing 3'-phosphotyrosine).
  • B. and C are representative gel showing inhibition of Tdpl activity by Compound A (NSC 88915) on three different substrates (i.e. single stranded, blunt duplex and 5 '-tailed duplex containing 3'-phosphotyrosine).
  • administration includes routes of introducing the compound(s) to a subject to perform their intended function.
  • routes of administration include injection (subcutaneous, intravenous, parenterally, intraperitoneally, intrathecal), oral, inhalation, rectal and transdermal.
  • admixture refers to something that is produced from mixing.
  • alkyl refers to the radical of saturated aliphatic groups, including straight- chain alkyl groups and branched-chain alkyl groups.
  • alkyl further includes alkyl groups, which can further include oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more carbons of the hydrocarbon backbone.
  • a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci-C 30 for straight chain, C 3 -C 3O for branched chain), preferably 26 or fewer, and more preferably 20 or fewer.
  • alkyl as used throughout the specification and claims is intended to include both “unsubstituted alkyls” and “substituted alkyls,” the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone.
  • alkyl also includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.
  • lower alkyl as used herein means an alkyl group, as defined above, but having from one to ten carbons, more preferably from one to six, and most preferably from one to four carbon atoms in its backbone structure, which may be straight or branched-chain.
  • alkylaryl or “aralkyl” are used interchangeably, and refer to an alkyl substituted with an aryl (e.g., phenylmethyl (benzyl)), or an aryl group substituted with an alkyl.
  • aryl e.g., phenylmethyl (benzyl)
  • heterooaralkyl refers to either an alkylaryl or aralkyl groups that is substituted at any number of positions with a heteroatom.
  • alkoxyalkyl refers to alkyl groups, as described above, which further include oxygen, nitrogen or sulfur atoms replacing one or more carbons of the hydrocarbon backbone, e.g., oxygen, nitrogen or sulfur atoms.
  • alkenyl and alkynyl refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
  • aryl refers to the radical of aryl groups, including 5- and 6- membered single-ring aromatic groups.
  • Heteroaryl groups may include from one to four heteroatoms. Examples of aryl and heteroaryl groups include benzene, pyrrole, furan, thiophene, imidazole, benzoxazole, benzothiazole, triazole, tetrazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
  • Polycyclic fused aromatic groups such as naphthyl, quinolyl, indolyl, and the like are also contemplated.
  • aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles," “heteroaryls” or “heteroaromatics.”
  • the aromatic ring can be substituted at one or more ring positions with such substiruents as described above, as for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino
  • cancer refers to a malignant tumor of potentially unlimited growth that expands locally by invasion and systemically by metastasis.
  • cancer also refers to the uncontrolled growth of abnormal cells. Specific cancers are selected from, but not limited to, rhabdomyosarcomas, chorio carcinomas, glioblastoma multiformas (brain tumors), bowel and gastric carcinomas, leukemias, ovarian cancers, prostate cancers, lymphomas, osteosarcomas or cancers which have metastasized.
  • carcinoma is art recognized and refers to malignancies of epithelial or endocrine tissues including respiratory system carcinomas, gastrointestinal system carcinomas, genitourinary system carcinomas, testicular carcinomas, breast carcinomas, prostatic carcinomas, endocrine system carcinomas, and melanomas.
  • Exemplary carcinomas include those forming from tissue of the cervix, lung, prostate, breast, head and neck, colon and ovary.
  • carcinosarcomas e.g., which include malignant tumors composed of carcinomatous and sarcomatous tissues.
  • An "adenocarcinoma” refers to a carcinoma derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • chiral refers to molecules which have the property of non-superimposability of the mirror image partner, while the term “achiral” refers to molecules which are superimposable on their mirror image partner.
  • cycloalkyl refers to the radical of saturated or unsaturated cyclic aliphatic groups, including cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.
  • cycloalkyl further includes cycloalkyl groups, which can further include oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more carbons of the hydrocarbon backbone.
  • Preferred cycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3, 4, 5, 6 or 7 carbons in the ring structure.
  • diastereomers refers to stereoisomers with two or more centers of dissymmetry and whose molecules are not mirror images of one another.
  • deuteroalkyl refers to alkyl groups in which one or more of the of the hydrogens has been replaced with deuterium.
  • an effective amount includes an amount effective, at dosages and for periods of time necessary, to achieve the desired result.
  • An effective amount of compound may vary according to factors such as the disease state, age, and weight of the subject, and the ability of the compound to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response.
  • An effective amount is also one in which any toxic or detrimental effects (e.g., side effects) of the angiogenesis inhibitor compound are outweighed by the therapeutically beneficial effects.
  • a therapeutically effective amount of compound may range from about 0.001 ⁇ g/kg/day to about 500 mg/kg/day, preferably 0.01 ⁇ g/kg/day and 100 mg/kg/day.
  • an effective dosage may range from about 0.001 ⁇ g/kg/day to about 500 mg/kg/day, preferably 0.01 ⁇ g/kg/day and 100 mg/kg/day.
  • certain factors may influence the dosage required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present.
  • treatment of a subject with a therapeutically effective amount of a compound can include a single treatment or, preferably, can include a series of treatments. It will also be appreciated that the effective dosage of a compound used for treatment may increase or decrease over the course of a particular treatment.
  • enantiomers refers to two stereoisomers of a compound which are non- superimposable mirror images of one another.
  • An equimolar mixture of two enantiomers is called a “racemic mixture” or a “racemate.”
  • halogen designates -F, -Cl, -Br or -I.
  • haloalkyl is intended to include alkyl groups as defined above that are mono-, di- or polysubstituted by halogen, e.g., fiuoromethyl and trifluoromethyl.
  • hydroxyl means -OH.
  • heteroatom as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.
  • heterocycloalkyl refers to the radical of saturated or unsaturated cyclic aliphatic groups substituted by any number of heteroatoms, including heterocycloalkyl (alicyclic) groups, alkyl substituted heterocycloalkyl groups, and heterocycloalkyl substituted alkyl groups.
  • Heteroatoms include but are not limited to oxygen, nitrogen, sulfur or phosphorous atoms replacing one or more carbons of the hydrocarbon backbone.
  • Preferred heterocycloalkyls have from 3-10 carbon atoms in their ring structure, and more preferably have 3, 4, 5, 6 or 7 carbons in the ring structure, wherein a heteroatom may replace a carbon atom.
  • hyperproliferative and neoplastic are used interchangeably, and include those cells having the capacity for autonomous growth, i.e., an abnormal state or condition characterized by rapidly proliferating cell growth.
  • Hyperproliferative and neoplastic disease states may be categorized as pathologic, i.e., characterizing or constituting a disease state, or may be categorized as non-pathologic, i.e., a deviation from normal but not associated with a disease state.
  • the term is meant to include all types of cancerous growths or oncogenic processes, metastatic tissues or malignantly transformed cells, tissues, or organs, irrespective of histopathologic type or stage of invasiveness.
  • “Pathologic hyperproliferative” cells occur in disease states characterized by malignant tumor growth. Examples of non-pathologic hyperproliferative cells include proliferation of cells associated with wound repair.
  • inhibitors refer to a method of prohibiting a specific action or function.
  • inhibitor refers to a molecule, compound or complex which blocks or modulates a biological or immunological activity.
  • isomers or “stereoisomers” refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.
  • leukemia is intended to have its clinical meaning, namely, a neoplastic disease in which white corpuscle maturation is arrested at a primitive stage of cell development.
  • the condition may be either acute or chronic.
  • Leukemias are further typically categorized as being either lymphocytic i.e., being characterized by cells which have properties in common with normal lymphocytes, or myelocytic (or myelogenous), i.e., characterized by cells having some characteristics of normal granulocytic cells.
  • Acute lymphocytic leukemia arises in lymphoid tissue, and ordinarily first manifests its presence in bone marrow.
  • AML Acute myelocytic leukemia
  • myeloblastic leukemia myeloblastic leukemia
  • promyelocytic leukemia myelomonocytic leukemia
  • myelomonocytic leukemia myelogenous leukemias as well.
  • leukemic cancer refers to all cancers or neoplasias of the hemopoietic and immune systems (blood and lymphatic system).
  • Chronic myelogenous leukemia also known as chronic granulocytic leukemia (CGL)
  • CML chronic myelogenous leukemia
  • CGL chronic granulocytic leukemia
  • modulate refers to increases or decreases in the activity of a cell in response to exposure to a compound of the invention, e.g., the inhibition of proliferation and/or induction of differentiation of at least a sub-population of cells in an animal such that a desired end result is achieved, e.g. , a therapeutic result, hi preferred embodiments, this phrase is intended to include hyperactive conditions that result in pathological disorders.
  • Neoplasia refers to "new cell growth” that results as a loss of responsiveness to normal growth controls, e.g. to neoplastic cell growth.
  • a “hyperplasia” refers to cells undergoing an abnormally high rate of growth.
  • the terms neoplasia and hyperplasia can be used interchangably, as their context will reveal, referring to generally to cells experiencing abnormal cell growth rates.
  • Neoplasias and hyperplasias include “tumors,” which may be either benign, premalignant or malignant.
  • non-direct interaction refers to any interactions that are not ionic nor covalent, such as hydrogen bonding or van der Waals interactions.
  • optionally substituted can include, for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl, sulfonamido, nitro,
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • a “peptide” is a sequence of at least two amino acids. Peptides can consist of short as well as long amino acid sequences, including proteins.
  • polycyclic group refers to the radical of two or more cyclic rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Rings that are joined through non-adjacent atoms are termed "bridged" rings.
  • Each of the rings of the polycycle can be substituted with such substituents as described above, as for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl,
  • prodrug includes compounds with moieties which can be metabolized in vivo. Generally, the prodrugs are metabolized in vivo by esterases or by other mechanisms to active drugs. Examples of prodrugs and their uses are well known in the art (See, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).
  • the prodrugs can be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compound in its free acid form or hydroxyl with a suitable esterifying agent. Hydroxyl groups can be converted into esters via treatment with a carboxylic acid.
  • prodrug moieties include substituted and unsubstituted, branch or unbranched lower alkyl ester moieties, ⁇ e.g. , propionoic acid esters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl esters (e.g., dimethylaminoethyl ester), acylamino lower alkyl esters (e.g., acetyloxymethyl ester), acyloxy lower alkyl esters (e.g., pivaloyloxymethyl ester), aryl esters (phenyl ester), aryl-lower alkyl esters (e.g., benzyl ester), substituted (e.g., with methyl, halo, or methoxy substituents) aryl and aryl-lower alkyl esters, amides, lower-alkyl amides, di-lower alkyl amides, and hydroxy amides.
  • protein refers to series of amino acid residues connected one to the other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. In general, the term “protein” is used to designate a series of greater than 50 amino acid residues connected one to the other.
  • reduced toxicity is intended to include a reduction in any undesired side effect elicited by a compound when administered in vivo.
  • the term “sarcoma” is art recognized and refers to malignant tumors of mesenchymal derivation.
  • subject refers to animals such as mammals, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like. In certain embodiments, the subject is a human.
  • sulfhydryl or "thiol” means -SH.
  • systemic administration means the administration of a compound(s), drug or other material, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • test compound or “drug candidate” or “modulator” or grammatical equivalents as used herein describes any molecule, e.g., protein, oligopeptide, small organic molecule, polysaccharide, polynucleotide, etc., to be tested for the capacity to directly or indirectly modulate the activity of Tdpl.
  • terapéuticaally effective amount refers to that amount of the compound being administered sufficient to prevent development of or alleviate to some extent one or more of the symptoms of the condition or disorder being treated.
  • treating and “treatment” refer to a method of alleviating or abating a disease and/or its attendant symptoms.
  • tumor suppressor gene refers to a gene that acts to suppress the uncontrolled growth of a cancer, such as a tumor.
  • tyrosine-DNA phosphodiesterase and “TDP” refer to a protein that is encoded by a tyrosine-DNA phosphodiesterase gene sequence or to a protein.
  • the terms refer to enzymes that cleave the phosphodiester bond linking the active site tyrosine residue of topoisomerase I with 3 '-terminus of DNA in topo I-DNA complexes.
  • the invention provides a method of inhibiting Tdpl activity in a subject, the method comprising administering to the subject a steroid compound capable of modulating the activity of Tdpl.
  • the invention provides a method of inhibiting Tdpl activity in a subject, the method comprising administering to the subject a compound of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(R b )N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R a , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR 3 , or NR b ; each R 3 is independently, H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, haloalkyl, -0R b , -SR b , -NR b R b , hydroxylalkyl, alkylcarbonylalkyl, mercaptoalkyl, aminoalkyl, sulfonylalkyl, sulfonylaryl, or thioalkoxy; each of which may be optionally substituted; each R b is independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, or heteroaryl; each of which may be optionally substituted; each m is independently 0, 1
  • the invention provides a method wherein X is
  • X is
  • R a is SCH 3 .
  • the invention provides a method wherein X is O-G-R. In a further embodiment, each G is S(O) 2 In another further embodiment, the invention provides a method wherein R is a substituted aryl.
  • the invention provides a method wherein the subject is identified as being in need of inhibition of Tdpl activity, and the compound of formula I is administered to the identified subject.
  • the invention provides a method of treating a Tdpl -related disorder in a subject, comprising administering to said subject in need thereof, an effective amount of a compound of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R 3 , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR a , or NR b ; each R a is independently, H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, haloalkyl, -0R b , -SR b , -NR b R b , hydroxylalkyl, alkylcarbonylalkyl, mercaptoalkyl, aminoalkyl, sulfonylalkyl, sulfonylaryl, or thioalkoxy; each of which may be optionally substituted; each R b is independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, or heteroaryl; each of which may be optionally substituted; each R
  • the invention provides a method wherein the disorder is cancer, tumor, neoplasm, neovascularization, vascularization, cardiovascular disease, intravasation, extravasation, metastasis, arthritis, infection, Alzheimer's Disease, blood clot, atherosclerosis, melanoma, skin disorder, rheumatoid arthritis, diabetic retinopathy, macular edema, or macular degeneration, inflammatory and arthritic disease, or osteosarcoma.
  • the disorder is cancer, tumor, neoplasm, neovascularization, vascularization, cardiovascular disease, intravasation, extravasation, metastasis, arthritis, infection, Alzheimer's Disease, blood clot, atherosclerosis, melanoma, skin disorder, rheumatoid arthritis, diabetic retinopathy, macular edema, or macular degeneration, inflammatory and arthritic disease, or osteosarcoma.
  • the invention provides a method of treating cancer in a subject identified as in need of such treatment, the method comprising administering to said subject an effective amount of a compound of formula I: I wherein,
  • A is a single or double bond
  • Y is H, OH, 0R b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R a , -OC(O)R a , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b S0 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(0)R a R a , each of which may be optionally substituted;
  • the invention provides a method wherein X is
  • X is
  • R a is SCH 3 .
  • the invention provides a method wherein X is O-G-R.
  • each G is S(O) 2.
  • R is a substituted aryl.
  • the invention provides a method wherein the compound is a Tdpl inhibitor.
  • the invention provides a method as described herein wherein the compound of formula I is:
  • the invention provides a method further comprising an additional therapeutic agent.
  • the additional therapeutic agent is an anticancer compound.
  • the invention provides a method wherein the step of administering the compound comprises administering the compound orally, topically, parentally, intravenously or intramuscularly. In another embodiment, the invention provides a method comprising the step of administering an effective amount of a composition comprising a compound of formula I and a pharmaceutically suitable excipient.
  • the invention provides a method, wherein the subject is a human.
  • the invention provides a method wherein the step of administering the compound of formula I comprises administering the compound in a dosage of between about 0.01 ⁇ g/kg/day and 100 mg/kg/day.
  • the invention provides for the use of a steroid compound in the manufacture of a medicament for inhibiting or reducing cancer in a patient, the compound being of formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R 3 )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkyl carbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R a , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR a , or NR b ; each R a is independently, H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, haloalkyl, -OR b , -SR b , -NR b R b , hydroxylalkyl, alkylcarbonylalkyl, mercaptoalkyl, aminoalkyl, sulfonylalkyl, sulfonylaryl, or thioalkoxy; each of which may be optionally substituted; each R b is independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, or heteroaryl; each of which may be optionally substituted; each of which
  • a subject is identified as being in need of Tdpl inhibition, hi certain embodiments, the compounds of the invention treat cancer in a subject by inhibiting Tdpl .
  • Tumors or neoplasms include new growths of tissue in which the multiplication of cells is uncontrolled and progressive. Some such growths are benign, but others are termed “malignant,” leading to death of the organism. Malignant neoplasms or “cancers” are distinguished from benign growths in that, in addition to exhibiting aggressive cellular proliferation, they invade surrounding tissues and metastasize. Moreover, malignant neoplasms are characterized in that they show a greater loss of differentiation (greater "dedifferentiation"), and of their organization relative to one another and their surrounding tissues. This property is also called “anaplasia.”
  • Neoplasms treatable by the present invention include all solid tumors, i.e., carcinomas and sarcomas, including Kaposi's sarcoma.
  • Carcinomas include those malignant neoplasms derived from epithelial cells which tend to infiltrate (invade) the surrounding tissues and give rise to metastases.
  • Adenocarcinomas are carcinomas derived from glandular tissue or in which the tumor cells form recognizable glandular structures.
  • Sarcoma, including Kaposi's sarcoma broadly include tumors whose cells are embedded in a fibrillar or homogeneous substance like embryonic connective tissue.
  • the invention is particularly illustrated herein in reference to treatment of certain types of experimentally defined cancers.
  • standard state-of-the-art in vitro and in vivo models have been used. These methods can be used to identify agents that can be expected to be efficacious in in vivo treatment regimens.
  • the method of the invention is not limited to the treatment of these tumor types, but extends to any solid tumor derived from any organ system. Cancers whose invasiveness or metastasis is associated with MMP expression, particularly gelatinase expression, are especially susceptible to being inhibited or even induced to regress by means of the invention.
  • the treatable cancers include, for example, colon cancer, bladder cancer, breast cancer, melanoma, ovarian carcinoma, prostatic carcinoma, or lung cancer, and a variety of other cancers as well.
  • the invention is especially useful in the inhibition of cancer growth in adenocarcinomas, including, for example, those of the prostate, breast, kidney, ovary, testes, and colon.
  • the invention is further useful against melanomas, which derive from the melanocyte system in the skin and other organs.
  • a solid tumor can be malignant, e.g. tending to metastasize and being life threatening, or benign.
  • solid tumors that can be treated according to a method of the present invention include sarcomas and carcinomas such as, but not limited to: fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas,
  • tumors comprising dysproliferative changes are treated or prevented in epithelial tissues such as those in the cervix, esophagus, and lung.
  • the present invention provides for treatment of conditions known or suspected of preceding progression to neoplasia or cancer, in particular, where non-neoplastic cell growth consisting of hyperplasia, metaplasia, or most particularly, dysplasia has occurred (for review of such abnormal growth conditions, see Robbins and Angell, 1976, Basic Pathology, 2d Ed., W. B. Saunders Co., Philadelphia, pp. 68-79).
  • Hyperplasia is a form of controlled cell proliferation involving an increase in cell number in a tissue or organ, without significant alteration in structure or function. As but one example, endometrial hyperplasia often precedes endometrial cancer. Metaplasia is a form of controlled cell growth in which one type of adult or fully differentiated cell substitutes for another type of adult cell. Metaplasia can occur in epithelial or connective tissue cells. Atypical metaplasia involves a somewhat disorderly metaplastic epithelium. Dysplasia is frequently a forerunner of cancer, and is found mainly in the epithelia; it is the most disorderly form of non-neoplastic cell growth, involving a loss in individual cell uniformity and in the architectural orientation of cells.
  • Dysplastic cells often have abnormally large, deeply stained nuclei, and exhibit pleomorphism. Dysplasia characteristically occurs where there exists chronic irritation or inflammation, and is often found in the cervix, respiratory passages, oral cavity, and gall bladder. For a review of such disorders, see Fishman et al., 1985, Medicine, 2d Ed., J. B. Lippincott Co., Philadelphia.
  • a method of the present invention may also be used to treat psoriasis, a dermatologic condition that is characterized by inflammation and vascular proliferation; benign prostatic hypertrophy, a condition associated with inflammation and possibly vascular proliferation; and cutaneous fungal infections. Treatment of other hyperproliferative disorders is also contemplated.
  • the present invention is directed to a method for inhibiting cancer growth, including processes of cellular proliferation, invasiveness, and metastasis in biological systems.
  • the method includes the use of a steroid compound as an inhibitor of cancer growth.
  • the method is employed to inhibit or reduce cancer cell proliferation, invasiveness, metastasis, or tumor incidence in living animals, such as mammals.
  • the invention includes a method of inducing cytotoxicity (cell killing) in cancer cells or reducing the viability of cancer cells.
  • the invention can be used to induce cytotoxicity in cells of carcinomas of the prostate, breast, ovary, testis, lung, colon, or breast.
  • the selective killing of the cancer cells can occur through apoptosis, necrosis, another mechanism, or a combination of mechanisms.
  • the killing of cancer cells can occur with less cytotoxicity to normal cells or tissues than is found with conventional cytotoxic therapeutics, preferably without substantial cytotoxicity to normal cells or tissues.
  • the steroid identified in the instant invention can induce cytotoxicity in cancer cells while producing little or substantially no cytotoxicity in normal cells.
  • the steroids can produce differential cytotoxicity: tumor cells are selectively killed whereas normal cells are spared.
  • the invention is a method for inducing differential cytotoxicity in cancer cells relative to normal cells or tissue. This differential in cytotoxicity associated with the steroid compounds occurs as a result of apoptosis, necrosis, another mechanism, or a combination of such mechanisms.
  • the steroid compounds exhibit their cancer treatment properties at concentrations that lead to fewer side effects than those of known chemotherapeutic agents, and in some cases are substantially free of side effects.
  • the steroid compounds are useful for extended treatment protocols, where other compounds would exhibit undesirable side-effects.
  • the properties of hydrophilicity and hydrophobicity are well balanced in these compounds, enhancing their utility both in vitro and especially in vivo, while other compounds lacking such balance are of substantially less utility.
  • the compounds have an appropriate degree of solubility in aqueous media to permit absorption and bioavailability in the body, while also having a degree of solubility in lipids to permit traversal of the cell membrane to a putative site of action. The compounds are maximally effective if they can be delivered to the site of the tumor and are able to enter the tumor cells.
  • the degree of hydrophilicity of the steroid compound can be of lesser importance.
  • the steroid compounds of the invention which may have low solubility in aqueous systems, can be used in direct or topical treatment of skin cancers, e.g., melanoma or basal cell carcinoma, or by implantation into the brain to topically treat brain cancer.
  • the steroid compounds are effective to inhibit the proliferation, invasiveness, or metastasis of cancer cells in vitro, as well as in vivo. These compounds possess an excellent balance of properties, in that they are shown to possess unusually strong activity in inhibiting the cancer growth, including proliferation, invasiveness, or metastasis of cancer cells. Another advantage is that it has an unexpectedly long serum half-life. Therefore, certain steroids may only require periodic administration, e.g., once or twice per week.
  • the method of the invention is effective to inhibit the enzymatic activity of matrix metalloproteinases, such as collagenases and gelatinases, associated with cancerous tumors in mammals.
  • matrix metalloproteinases such as collagenases and gelatinases
  • the gelatinolytic activity capable of inhibition may derive from gelatinase expression by the cancerous tumor or from normal, i.e., non-cancerous, tissue.
  • the gelatinase activity may be derived from such normal tissues as epithelial tissue or stromal tissue. More preferably, the method can be used to inhibit excessive gelatinolytic activity associated with such tumors.
  • This inhibition of observed gelatinolytic activity may be due to inhibition of MMP activity, down-regulation of MMP expression, or some other interference with the physiology associated with these gelatinases, such as inhibition of activation of the precursor form of the enzyme, pro-gelatinase (or pro-MMP).
  • the inhibition of cancer cells may result from inhibition of MMP activity, down- regulation of MMP expression, some other mechanism, or a combination of mechanisms. It is believed that all solid cancer types that express MMPs or that exhibit invasive or metastatic properties can be treated by the method of the invention. In some cases, the incidence or development of tumor foci can be inhibited or substantially prevented from occurring. Therefore, the method can be used as a prophylactic treatment, e.g., by administering the steroid compound to a mammal after detection of a gene product or metabolite associated with predisposition to a cancer but before any specific cancerous lesion is detected. Alternatively, the steroid compounds are useful for preventing cancer recurrence, for example, to treat residual cancer following surgical resection or radiation therapy.
  • the effect occurs over a wide range of concentrations, including at concentrations that are extraordinarily low.
  • the amount of the steroid compound used according to the invention is an amount that is effectively inhibitory of cancer growth.
  • An amount of a steroid compound is effectively inhibitory to cancer growth if it significantly reduces cellular proliferation or the potential of invasiveness or metastasis.
  • Proliferation refers to the capacity of a tumor to increase its volume through cell division, typically measured as the "doubling rate.”
  • the inhibition of cellular proliferation by the present method means that the rate of growth is decreased, hi some cases, the method can actually induce regression or diminution of tumor mass, if the rate of replenishment of the tumor cells through cell division is exceeded by the rate of cell death.
  • Invasiveness refers to the potential of a tumor or tumor cells to invade other tissues, typically by breaking down the extracellular matrix of those tissues.
  • Metastasis refers to the potential of a tumor or tumor cells to establish new tumor foci at sites distant from the primary site where the tumor began.
  • metastasis proceeds by individual cells or groups of cells breaking off from the primary tumor and migrating, e.g., through the blood or lymph, to establish a new tumor focus in another tissue or organ.
  • One locus common in tumor metastasis is in the lung, where the very fine vasculature of the lung tissue can often catch circulating tumor cells, permitting the establishment of a tumor focus therein.
  • the cancers treatable by means of the present invention occur in mammals.
  • Mammals include, for example, humans, as well as pet animals such as dogs and cats, laboratory animals such as rats and mice, and farm animals such as horses and cows.
  • Tdpl is a member of the phospholipase D superfamily, which encompasses a diverse group of enzymes that catalyze phosphodiester bond cleavage on substrates ranging from phospholipids to DNA, or in the case of Tdpl , protein-DNA complexes.
  • the majority of the sequence identity between Tdpl and the members of the PLD family is restricted to two copies of an active site signature motif H(X)K(X 4 )D, also referred to as an HKD motif, which has been implicated in the catalytic mechanism of these enzymes.
  • Tdpl represents a distinct subclass of the PLD superfamily based on its unique "HKN" motifs.
  • the importance of the two signature HKN motifs for Tdpl enzymatic activity has been established by site-directed mutagenesis.
  • Tdpl The affiliation of Tdpl with the PLD superfamily has been further established through comparative analysis of the three-dimensional structure of the catalytically active N-terminal truncated human Tdpl ( ⁇ l-148) (Fig. IA) (Davies, D.R., Interthal, H., Champoux, J.J., and HoI, W. G. Structure, 2002, 10. 237) with the known crystal structures of two members of the PLD superfamily, a PLD from Streptomyces and a bacterial nuclease from Salmonella tryphimurium. Albeit the low sequence identity amid these three enzymes, they share a strikingly similar tertiary structure.
  • Tdpl crystal structure also reveals that four additional residues, which are conserved in all Tdpl orthologs, N283, Q294, N516, and E538, are also positioned near the active site (Davies, D.R., Interthal, H., Champoux, J.J., and HoI, W.G. Structure, 2002, 10. 237).
  • Fig. 1C The molecular assembly of vanadate, Tdpl, a Topi -derived peptide containing the active site tyrosine, and a single-stranded DNA oligonucleotide into a quaternary structure is shown in Fig. 1C.
  • Initial examination of the quaternary structure revealed an asymmetrical substrate-binding channel that extended in opposite directions from the active site across the entire surface of Tdpl.
  • the DNA moiety bound to one side of the active site in a relatively narrow cleft that is predominantly positively charged.
  • the peptide moiety bound to the other side of the active site in a relatively large, more open cleft that contains a mixed charge distribution.
  • a closer look into the active site of the quaternary structure shows that the vanadate ion exhibits trigonal bipyramidal geometry, which is consistent with a transition state of an S N 2 nucleophilic attack on phosphate.
  • the apical coordination to the vanadate molecule is filled by the imidazole N ⁇ 2 atom of the active site H263 residue of Tdpl and the oxygen atom of the tyrosine side chain of the Topi -derived peptide, whereas the 3'-hydroxyl of the DNA oligonucleotide occupies one of the three equatorial positions.
  • the H493 residue from the C- terminal HKN motif acts as a general acid and donates a proton to the apical tyrosine-containing peptide-leaving group (Fig. 2A). This results in the formation of a transient covalent phosphoamide bond between the N ⁇ 2 atom of H263 and the 3'-end of the DNA (Fig. 3B). Hydrolysis of this covalent intermediate is proposed to be carried out by a water molecule that is activated by the H493 residue acting as a general base (Fig. 2C).
  • the second S N 2 reaction step in the Tdpl catalytic mechanism is supported by in vitro biochemical studies with the SCANl H493R mutant, which leads to an accumulation of Tdpl -DNA covalent intermediate.
  • the final product released after Tdpl catalysis is a DNA molecule with a 3 '-phosphate end (Fig. 2D).
  • a stalled Topi -DNA complex is an unmanageable substrate for Tdpl because of the limited access of the scissile phosphotyrosyl bond, which is internally located within the Topl- DNA complex (Redinbo, M. R., Stewart, L., Kuhn, P., Champoux, J.J., and HoI, W. G. Science, 1998, 279. 1504).
  • This statement is corroborated by the poor processing by Tdpl of a suicide complex made of full-length human Topi covalently linked to a DNA oligonucleotide (Fig. 3F (iii)) (Debethune, L., Kohlhagen, G., Grandas, A., and Pommier, Y.
  • Tdpl substrates Besides the physiological substrates, numerous synthetic Tdpl substrates have been identified (Fig. 3D). Specifically, phosphotyrosine analogues, such as 3'-(4-nitro)phenol and 3'(4-methyl)phenol, have been used in kinetic analysis of the Tdpl catalytic mechanism.
  • Other artificial substrates include a 3'-linker-biotin (Interthal, H., Chen, HJ., and Champoux, JJ. J Biol Chem, 2005, 280. 36518) and a fluorescent 3'-4-methylumbelliferone (Nitiss, K.C., Malik, M., He, X., White, S.W., and Nitiss, J.L. Proc Natl Acad Sci USA, 2006, 103. 8953), which could potentially be employed in high-throughput screens for Tdpl inhibitors, similar to how the 3'-BV tag has been used (discussed below).
  • Tdpl The 3 '-phosphate-end generated by Tdpl must be hydrolyzed to a 3'-hydroxyl in order for further DNA repair to occur.
  • Polynucleotide kinase phosphatase (PNKP) a bifunctional enzyme with 5 '-kinase and 3 '-phosphatase activities, has been suggested as a reasonable candidate in human cells for the repair of these 3 '-phosphate lesions.
  • T4 kinase the bacteriophage equivalent of PNKP
  • Tdpl Huizenga, B.N., Robertson, C.A., Yao, K.C, and Nash, YL. A. Proc Natl Acad Sci USA, 1996, 93. 11534
  • PNKP is known to interact with the XRCCl protein, which together with the other base excision repair (BER) proteins, such as DNA polymerase ⁇ , DNA ligase III, and poly(ADP-ribose)polym erase 1(P ARP-I) forms a multiprotein DNA repair complex (Whitehouse, CJ. , Taylor, R.M., Thistlethwaite, A., Zhang, H., Karimi-Busheri, F., Lasko, D. D., Weinfeld, M., and Caldecott, K. W. Cell, 2001, 104. 107).
  • BER base excision repair
  • Tdpl has also been shown to be a member of the BER repair complex through interactions with DNA ligase III (El-Khamisy, S. F., Saifi, G.M., Weinfeld, M., Johansson, F., Helleday, T., Lupski, J.R., and Caldecott, K.W. Nature, 2005, 434. 108) and XRCCl (PIo, L, Liao, Z. Y., Barcelo, J.M., Kohlhagen, G., Caldecott, K.W., Weinfeld, M., and Pommier, Y. DNA Repair (Amst), 2003, 2. 1087).
  • DNA ligase III El-Khamisy, S. F., Saifi, G.M., Weinfeld, M., Johansson, F., Helleday, T., Lupski, J.R., and Caldecott, K.W. Nature, 2005, 434. 108
  • XRCCl
  • XRCCl -deficient cells have been discovered to be hypersensitive to CPT and defective in Tdpl activity, which is in agreement with a direct role of Tdpl in association with the BER complex for the removal and repair of Topi -mediated DNA damage.
  • the interaction of XRCCl with PNKP has been shown to stimulate both the 5 '-kinase and 3 '-phosphatase activities of this enzyme (Whitehouse, C. J., Taylor, R.M., Thistlethwaite, A., Zhang, H., Karimi-Busheri, F., Lasko, D. D., Weinfeld, M., and Caldecott, K.W. Cell, 2001, 104. 107).
  • XRCCl interacts with DNA ligase III and increases the intracellular stability of the ligase (Caldecott, K.W., Tucker, J. D., Stanker, L.H., and Thompson, L.H. Nucleic Acids Res, 1995, 23. 4836; Taylor, R.M., Wickstead, B., Cronin, S., and Caldecott, K.W. Cur r Biol, 1998, 8. 877).
  • the association PARP-I with the XRCCl complex has also been observed. However, its function in the repair of Topi -mediated DNA damage remains to be determined.
  • PARP-I is a nuclear enzyme often termed a molecular nick sensor, which recognizes and binds to DNA single or double strand breaks.
  • Tdpl activity after the association with the XRCCl complex is still unresolved.
  • Tdpl hydrolyzes the Topl-DNA bond.
  • PNKP hydrolyzes the resulting 3'-phosphate end and catalyzes the phosphorylation of the 5 '-end of the DNA.
  • DNA polymerase ⁇ replaces the missing DNA segment and DNA ligase III reseals the DNA.
  • Tdpl is one of several potentially redundant pathways involved in the repair of Top-mediated damage in yeast (Pouliot, J.J., Robertson, C.A., and Nash, H.A. Genes Cells, 2001, 6.
  • RadllRadlO functions primarily in the nucleotide excision repair pathway, where it cleaves on the 5 '-side of the repair bubble formed around bulky DNA lesions (Vance, J.R., and Wilson, T.E. Proc Natl Acad Sci USA, 2002, 99. 13669; Roberts, J.A., and White, M.F. Nucleic Acids Res, 2005, 33. 6662; Bardwell, A.J., Bardwell, L., Tomkinson, A.E., and Friedberg, E.C. Science, 1994, 265. 2082).
  • Tdpl inhibition Even though there is redundancy in the repair of Topi -mediated DNA damage, it may not be critical to the clinical relevance of Tdpl inhibition, since it is well established that deficiencies in checkpoint mechanisms are a common feature in cancer cells (Dasika, G. K., Lin, S. C, Zhao, S., Sung, P., Tomkinson, A., and Lee, E. Y. Oncogene, 1999, 18. 7883). For example, if cancer-related checkpoint inactivation arises, then Tdpl becomes the principle source for the removal of Topi -mediated DNA damage (Fig. 4B).
  • Topi DNA topoisomerase I
  • Topi inhibitors damage DNA by trapping covalent complexes between the Topi catalytic tyrosine and the 3 '-end of the broken DNA.
  • Tyrosyl-DNA phosphodiesterase (Tdpl) repairs Topi -DNA covalent complexes by hydrolyzing the tyrosyl-DNA bond.
  • Tdpl inhibitors are therefore useful as anticancer agents both in monotherapy and in combination with other anticancer compounds (particularly DNA-targeted anticancer compounds) such as Topi inhibitors.
  • Tumor cells whose repair pathways are commonly deficient, might be selectively sensitized to Topi inhibitors compared to normal cells that contain redundant repair pathways.
  • Tdpl inhibitors are effective by themselves as anticancer agents as oncogenic activation tends to increase free radical production and genomic instability (Cerutti PA (1985) Science 227 (4685):375-381 ; Kc S et al. Mutat Res. (2006) 29 593(l-2):64-79.; Vafa et al., MoI Cell 9(5): 1031-1044 (2002)).
  • the invention provides methods for treating cancer and other cell proliferative disorders by administering to a subject in need thereof an effective amount of a combination of a Tdpl inhibitor of this invention together with a Topi inhibitor.
  • Topi inhibitors A variety of Topi inhibitors have been reported, including camptothecin, irinotecan, topotecan, saintopin, and derivatives and analogs thereof.
  • the invention provides pharmaceutical compositions including a Tdpl inhibitor of this invention together with a Topi inhibitor, optionally including a pharmaceutically-acceptable carrier or excipient.
  • the invention provides methods and compositions for the treatment or prevention of parasitic disease.
  • Tdpl inhibitors may be valuable as anti-infectious agents since the gene is present in parasites, including Trypanosoma brucei rhodesiense, Trypanosoma brucei gambiense, and Plasmodium spp. including P. vivax, P. falciparum, P. ovale, and P. malaria.
  • the invention provides methods for treating or preventing a parasitic infection caused by a parasite expressing Tbpl , the method including the step of administereing to a subject in need thereof an effective amount of a Tdpl inhibitor according to this invention.
  • the invention provides pharmaceutical compositions for treatment or preention of parasitic disease, including a Tdpl inhibitor of this invention together pharmaceutically- acceptable carrier or excipient.
  • the invention provides a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R ⁇ -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R 3 , -NR b S0 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • each W is independently C, N, CR a , or NR b ; each R a is independently, H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, haloalkyl, -OR b , -SR b , -NR b R b , hydroxylalkyl, alkylcarbonylalkyl, mercaptoalkyl, aminoalkyl, sulfonylalkyl, sulfonylaryl, or thioalkoxy; each of which may be optionally substituted; each R b is independently H, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, or heteroaryl; each of which may be optionally substituted; each of which
  • Tdpl A library that consists of 1981 compounds from the Developmental Therapeutics Program of the National Cancer Institute was screened against human Tdpl. These compounds are representative of the chemical and structural diversity of the available repository of more that 14,000 chemicals. Several inhibitors of Tdpl were identified. The steroid compounds as described above were discovered using this high- throughput screen and were confirmed by biochemical assays to be potent inhibitors of Tdpl with IC50s in the micromolar range. Specifically, a high-throughput screening assay to discover specific inhibitors of Tdpl using a novel electrochemiluminescence method has been utilized by the invention.
  • BV-TagTM ruthenium labels
  • Fig 5 upper panel ruthenium labels
  • BioVeris® M8 analyzer BioVeris Corp., Gaithersburg, MD
  • Tdpl DNA substrate to be used in the high throughput BioVeris assay was designed.
  • the invention utilized a 5'-biotinylated 14-mer single strand DNA substrate using the Tdpl DNA substrate sequence published previously (Z. Liao, L. Thibaut, A. Jobson et al., MoI Pharmacol 70 (1), 366 (2006).
  • This substrate consists in the 3' incorporation of a phosphotyrosine group which is an available DNA modification proposed by Glen Research (Sterling, VA).
  • This invention attached a BV-TagTM on the phosphotyrosyl free amine using a NHS ester BV-TagTM available from BioVeris Corp. This coupling reaction is straightforward and does not require any specific chemistry equipment. (Fig. 6).
  • Tdpl BV substrate gives a maximum signal in the instrument.
  • the substrate is processed, leading to the removal of the tyrosyl-BV- Tag group and therefore to a loss of signal (Fig. 7).
  • Positive hits highlight potential specific Tdpl inhibitors and maintain the electrochemiluminescence signal to its maximum level.
  • the 3000-compound plated chemical libraries from the NCl Developmental Therapeutic Program (DTP) was screened and identified the steroid NSC 88915 as a positive hit for Tdpl inhibition from the 1981 -compound Diversity Set (Fig. 9).
  • NSC 88915 and other derivatives were confirmed as Tdpl inhibitors in gel-based assays (Fig. 10).
  • Formulation, Administration, Kits, and Pharmaceutical Compositions were confirmed as Tdpl inhibitors in gel-based assays (Fig. 10).
  • the invention also provides a pharmaceutical composition, comprising an effective amount a compound described herein and a pharmaceutically acceptable carrier.
  • compound is administered to the subject using a pharmaceutically-acceptable formulation, e.g., a pharmaceutically-acceptable formulation that provides sustained delivery of the compound to a subject for at least 12 hours, 24 hours, 36 hours, 48 hours, one week, two weeks, three weeks, or four weeks after the pharmaceutically-acceptable formulation is administered to the subject.
  • the invention provides for a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R* 5 , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -OR b , -SR b , -NR a R a , -C(O)R 3 , -OC(O)R 3 , NR b C(O)R a , -C(NR)R 3 , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R a R a , each of which may be optionally substituted;
  • the invention provides for a kit comprising an effective amount of a steroid compound in unit dosage form, together with instructions for administering the steroid compound to a subject suffering from cancer, wherein the compound is of formula I: a compound of Formula I:
  • A is a single or double bond
  • Y is H, OH, OR b , S(O)R b , S(O) 2 R b , N(R b ) 2 , C(O)R b , C(S)R b , C(NR)N(R b ) 2 , C(O)N(R b ) 2 , (C(R a )(R b )) m N(R b ) 2 , or (C(R a )(R b )) m OR b ;
  • X is O-G-R; wherein G is S, S(O), or S(O) 2 ;
  • R is H, alkyl, aryl, aralkyl, heteroaryl, halo, haloalkyl, hydroxyalkyl, aminoalkyl, mercaptoalkyl, alkylcarbonyl, cycloalkyl, heterocycloalkyl, heteroaralkyl, -0R b , -SR b , -NR a R a , -C(O)R a , -OC(O)R 3 , NR b C(O)R a , -C(NR)R a , -OC(NR)R 3 , -NR b C(NR)R a , -NR b SO 2 R a , -OS(O) 2 R 3 , -OP(O)R 3 R 3 , or -P(O)R 3 R 3 , each of which may be optionally substituted;
  • phrases "pharmaceutically acceptable” refers to those compounds of the present invention, compositions containing such compounds, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically-acceptable carrier includes pharmaceutically-acceptable material, composition or vehicle, involved in carrying or transporting the subject chemical from one organ, or portion of the body, to another organ, or portion of the body.
  • Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.
  • a therapeutically effective amount can be administered in one or more doses.
  • administration or “administering” includes routes of introducing the compound(s) to a subject to perform their intended function. Examples of routes of administration which can be used include injection (subcutaneous, intravenous, parenterally, intraperitoneally, intrathecal), oral, inhalation, rectal and transdermal.
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • systemic administration means the administration of a compound(s), drug or other material, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • compositions include the step of bringing into association a compound(s) with the carrier and, optionally, one or more accessory ingredients.
  • These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.
  • a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
  • the compound(s), which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention are formulated into pharmaceutically-acceptable dosage forms by conventional methods known to those of skill in the art.
  • the pharmaceutical compositions are suitable for topical, intravenous, intratumoral, parental, or oral administration.
  • the methods of the invention further include administering to a subject a therapeutically effective amount of a conjugate in combination with another pharmaceutically active compound.
  • Pharmaceutically active compounds that may be used can be found in Harrison's Principles of Internal Medicine, Thirteenth Edition, Eds. T.R. Harrison et al. McGraw-Hill N. Y., NY; and the Physicians Desk Reference 50th Edition 1997, Oradell New Jersey, Medical Economics Co., the complete contents of which are expressly incorporated herein by reference.
  • Formulations are provided to a subject in an effective amount.
  • the term "effective amount" includes an amount effective, at dosages and for periods of time necessary, to achieve the desired result.
  • An effective amount of conjugate may vary according to factors such as the disease state, age, and weight of the subject, and the ability of the compound to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response. The effective amount is generally determined by the physician on a case-by-case basis and is within the skill of one in the art. As a rule, the dosage for in vivo therapeutics or diagnostics will vary. Several factors are typically taken into account when determining an appropriate dosage. These factors include age, sex and weight of the patient, the condition being treated, and the severity of the condition.
  • Suitable dosages and formulations of immune modulators can be empirically determined by the administering physician. Standard texts, such as Remington: The Science and Practice of Pharmacy, 17th edition, Mack Publishing Company, and the Physician's Desk Reference, each of which are incorporated herein by reference, can be consulted to prepare suitable compositions and doses for administration. A determination of the appropriate dosage is within the skill of one in the art given the parameters for use described herein. Standard texts, such as Remington: The Science and Practice of Pharmacy, 17th edition, Mack Publishing Company, incorporated herein by reference, can be consulted to prepare suitable compositions and formulations for administration, without undue experimentation. Suitable dosages can also be based upon the text and documents cited herein. A determination of the appropriate dosages is within the skill of one in the art given the parameters herein.
  • an effective amount is an amount that is sufficient to palliate, ameliorate, stabilize, reverse or slow the progression of a cancerous disease or otherwise reduce the pathological consequences of the cancer.
  • a therapeutically effective amount can be provided in one or a series of administrations. The effective amount is generally determined by the physician on a case-by-case basis and is within the skill of one in the art.
  • the dosage for in vivo therapeutics or diagnostics will vary. Several factors are typically taken into account when determining an appropriate dosage. These factors include age, sex and weight of the patient, the condition being treated, the severity of the condition and the form of the compound being administered.
  • the dosage of conjugates can range from about 0.001 ⁇ g/kg/day to about 500 mg/kg/day.
  • Methods for administering compositions are known in the art. Such dosages may vary, for example, depending on whether multiple administrations are given, tissue type and route of administration, the condition of the individual, the desired objective and other factors known to those of skill in the art. Administrations can be conducted infrequently, or on a regular weekly basis until a desired, measurable parameter is detected, such as diminution of disease symptoms. Administration can then be diminished, such as to a biweekly or monthly basis, as appropriate.
  • Such dosages may vary, for example, depending on whether multiple administrations are given, tissue type and route of administration, the condition of the individual, the desired objective and other factors known to those of skill in the art.
  • the composition Following administration of the composition, it can be necessary to wait for the composition to reach an effective tissue concentration at the site of the disorder before detection. Duration of the waiting step varies, depending on factors such as route of administration, location, and speed of movement in the body. In addition, where the compositions are coupled to molecular carriers, the rate of uptake can vary, depending on the level of receptor expression on the surface of the cells. For example, where there is a high level of receptor expression, the rate of binding and uptake is increased. Determining a useful range of waiting step duration is within the level of ordinary skill in the art and may be optimized.
  • the steroid compounds useful according to the method of the invention appear to exhibit their beneficial effect in a dose-dependent manner.
  • administration of larger quantities of a steroid compound has been observed to inhibit cancer cell growth or invasiveness to a greater degree than does administration of a smaller amount.
  • efficacy has been observed at dosages below the level at which toxicity is seen in normal cells or at the organismal level. Accordingly, one of the advantages of the invention is that the debilitating side effects usually attendant upon conventional cytotoxic cancer treatments are reduced, and preferably avoided.
  • Available routes of administration include subcutaneous, intramuscular, intraperitoneal, intradermal, oral, intranasal, intrapulmonary (i.e., by aerosol), intravenously, intramuscularly, subcutaneously, intracavity, intrathecally or transdermally, alone or in combination with other pharmaceutical agents.
  • compositions for oral, intranasal, or topical administration can be supplied in solid, semisolid or liquid forms, including tablets, capsules, powders, liquids, and suspensions.
  • Compositions for injection can be supplied as liquid solutions or suspensions, as emulsions, or as solid forms suitable for dissolution or suspension in liquid prior to injection.
  • a preferred composition is one that provides a solid, powder, or liquid aerosol when used with an appropriate aerosolizer device.
  • compositions are preferably supplied in unit dosage form suitable for administration of a precise amount.
  • Also contemplated by this invention are slow-release or sustained release forms, whereby a relatively consistent level of the active compound are provided over an extended period.
  • Another method of administration is intravascular, for instance by direct injection into the blood vessel, or surrounding area. Further, it may be desirable to administer the compositions locally to the area in need of treatment; this can be achieved, for example, by local infusion during surgery, by injection, by means of a catheter, or by means of an implant, said implant being of a porous, non-porous, or gelatinous material, including membranes, such as silastic membranes, or fibers.
  • Enteral administration is a preferred route of delivery of the steroid, and compositions including the steroid compound with appropriate diluents, carriers, and the like are readily formulated. Liquid or solid (e.g., tablets, gelatin capsules) formulations can be employed. It is among the advantages of the invention that, in many situations, the steroid compound can be delivered orally, as opposed to parenteral delivery (e.g., injection, infusion) which is typically required with conventional chemotherapeutic agents.
  • parenteral delivery e.g., injection, infusion
  • Parenteral use e.g., intravenous, intramuscular, subcutaneous injection
  • formulations using conventional diluents, carriers, etc., such as are known in the art can be employed to deliver the compound.
  • delivery of the steroid compound can include topical application.
  • Compositions deemed to be suited for such topical use include as gels, salves, lotions, ointments and the like.
  • the steroid compound can be delivered via a slow-release delivery vehicle, e.g., a polymeric material, surgically implanted at or near the lesion situs.
  • the maximal dosage for a subject is the highest dosage that does not cause undesirable or intolerable side effects.
  • side effects may include clinically significant antimicrobial or antibacterial activity, as well as toxic effects.
  • the practitioner is guided by skill and knowledge in the field, and the present invention includes, without limitation, dosages that are effective to achieve the described phenomena.
  • the invention can also be practiced by including with the steroid compound one or more other anti-cancer chemotherapeutic agents, such as any conventional chemotherapeutic agent.
  • chemotherapeutic agents such as any conventional chemotherapeutic agent.
  • the combination of the steroid compound with such other agents can potentiate the chemotherapeutic protocol.
  • Numerous chemotherapeutic protocols will present themselves in the mind of the skilled practitioner as being capable of incorporation into the method of the invention. Any chemotherapeutic agent can be used, including alkylating agents, antimetabolites, hormones and antagonists, radioisotopes, as well as natural products.
  • steroid compound can be administered with antibiotics such as doxorubicin and other anthracycline analogs, nitrogen mustards such as cyclophosphamide, pyrimidine analogs such as 5- fiuorouracil, cisplatin, hydroxyurea, taxol and its natural and synthetic derivatives, and the like.
  • antibiotics such as doxorubicin and other anthracycline analogs
  • nitrogen mustards such as cyclophosphamide
  • pyrimidine analogs such as 5- fiuorouracil, cisplatin, hydroxyurea, taxol and its natural and synthetic derivatives, and the like.
  • the steroid in the case of mixed tumors, such as adenocarcinomas of the breast and prostate, in which the tumors can include gonadotropin-dependent and gonadotropin-independent cells
  • the steroid can be administered in conjunction with leuprolide or goserelin.
  • antineoplastic protocols include the use of a steroid compound with another treatment modality, e.g., surgery, radiation, other chemotherapeutic agent, etc., referred to herein as "adjunct antineoplastic modalities.”
  • another treatment modality e.g., surgery, radiation, other chemotherapeutic agent, etc.
  • the method of the invention can be employed with such conventional regimens with the benefit of reducing side effects and enhancing efficacy.
  • Another embodiment is a compound of any of the formulae herein made by a process delineated herein, including the processes exemplified in the schemes and examples herein.
  • Another aspect of the invention is a compound of any of the formulae herein for use in the treatment or prevention in a subject of a disease, disorder or symptom thereof delineated herein.
  • Another aspect of the invention is use of a compound of any of the formulae herein in the manufacture of a medicament for treatment or prevention in a subject of a disease, disorder or symptom thereof delineated herein. Examples
  • Tdpl substrates HPLC purified oligonucleotides N14Y (PIo et al., (2003) DNA Repair (Amst) 2(10): 1087-1100) were labeled at their 5'-end with [ ⁇ - 32 P]-ATP (Perkin- Elmer Life Science Co., Boston, MA) by incubation with 3 '-phosphatase free T4 polynucleotide kinase (Roche applied Science, Indianapolis, IN) according to the manufacturer's protocols. Unincorporated nucleotides were removed by Sephadex G-25 spin-column chromatography (Mini Quick Spin Oligo Columns, Roche, Indianapolis, IN).
  • N14Y was mixed with the complementary oligonucleotide in equal molar ratios in annealing buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 10 mM MgCl 2 ), heated to 96 0 C, and allowed to cool down slowly (over 2 h) to room temperature.
  • Tdpl assays were performed in 20 ⁇ l mixtures containing 50 mM Tris-HCl, pH 8.0, 80 mM KCl, 2 mM EDTA, 1 mM dithiothreitol (DTT), and 40 ⁇ g/ml bovine serum albumin (BSA).
  • BSA bovine serum albumin
  • 25 nM of 5'- 32 P-labeled ' substrate (D 14Y) was reacted with 1 ng Tdpl ( ⁇ 0-7 nM) in the absence or presence of inhibitor for 20 min at 25°C.
  • Tdpl inhibitors have become a major area of drug research and structure-based design, with Tdpl, works synergistically and selectively in the cancer cells.
  • Tdpl can repair DNA topoisomerase I (Topi) covalent complexes by hydrolyzing the tyrosyl-DNA phosphodiester bond.
  • the natural substrate of Tdpl is large and complex, consisting of tyrosine or possibly a tyrosine-containing peptide moiety linked to a single strand of DNA via a 3' phosphodiester bond (Interthal, H.; Pouliot, J.J. PNAS, 98, 21 (2001)).
  • Example 1 The results obtained in Example 1 are used to refine a model for prediction of binding affinity of compounds against Tdpl as follows.
  • the obtained compounds will then be subjected to flexible docking and the compounds are selected based on the docking score for the Energy of the Model.
  • the results are compared with a training set of compounds found to bind to the active site of Tdpl .
  • Figure 1 IA is a representative gel showing inhibition of Tdpl activity by Compound A (NSC 88915) on three different substrates (i.e. single stranded, blunt duplex and 5'-tailed duplex containing 3 '-phospho tyrosine).
  • Figure B and C show inhibition by SCAN-I Tdpl mutant (H493R) cleavage activity and protein-DNA complex formation by Compound A (NSC 88915) using SDS-PAGE (B) and denaturing PAGE (C) gel electrophoresis.
  • SCAN-I Tdpl mutant H493R
  • B SDS-PAGE
  • C denaturing PAGE

Landscapes

  • Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne des composés stéroïdes et des compositions, ainsi que des procédés d'inhibition de l'activité Tdpl. L'invention concerne en outre des procédés de traitement de troubles associés au Tdpl, incluant le cancer, au moyen de dérivés de stéroïdes.
PCT/US2008/004541 2007-04-05 2008-04-05 Composés stéroïdes, compositions et procédés de traitement WO2008124145A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US92198007P 2007-04-05 2007-04-05
US60/921,980 2007-04-05
US43007P 2007-10-24 2007-10-24
US61/000,430 2007-10-24

Publications (1)

Publication Number Publication Date
WO2008124145A1 true WO2008124145A1 (fr) 2008-10-16

Family

ID=39683646

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/004541 WO2008124145A1 (fr) 2007-04-05 2008-04-05 Composés stéroïdes, compositions et procédés de traitement

Country Status (1)

Country Link
WO (1) WO2008124145A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8716295B2 (en) 2010-10-27 2014-05-06 Yves Pommier Fluoroquinolone derivatives or sulfonamide moiety-containing compounds as inhibitors of tyrosyl-dnaphosphodiesterase (TDP1)
US11691963B2 (en) 2020-05-06 2023-07-04 Ajax Therapeutics, Inc. 6-heteroaryloxy benzimidazoles and azabenzimidazoles as JAK2 inhibitors
US11970494B2 (en) 2021-11-09 2024-04-30 Ajax Therapeutics, Inc. 6-heteroaryloxy benzimidazoles and azabenzimidazoles as JAK2 inhibitors

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997024367A1 (fr) * 1995-12-29 1997-07-10 Glaxo Group Limited Derives de 21-(2-oxo-tetrahydrofurane)-thiopregnane, leur procede de preparation et compositions pharmaceutiques les contenant
WO2001072293A2 (fr) * 2000-03-24 2001-10-04 Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Antiprogestins a activite partielle d'agonistes
WO2003014139A1 (fr) * 2001-08-08 2003-02-20 San Diego State University Foundation Compositions therapeutiques et procedes de criblage lies a la 3'-tyrosyl-adn-phosphodiesterase
WO2003015757A1 (fr) * 2001-08-16 2003-02-27 The Trustees Of The University Of Pennsylvania Synthese et utilisation de reactifs pour ameliorer la lipofection d'adn et/ou les therapies par medicaments et promedicaments a liberation lente
WO2006015369A2 (fr) * 2004-08-03 2006-02-09 The Johns Hopkins University Dosage d'identification d'inhibiteurs de topoisomerase
WO2006099095A2 (fr) * 2005-03-11 2006-09-21 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Inhibiteurs de ribosome utilises en tant qu'inhibiteurs de la tyrosyl-dna-phosphodiesterase
WO2007112121A2 (fr) * 2006-03-27 2007-10-04 Government Of The United States Of Americas, As Represented By The Secretary Composés à base de tétracyline et procédés de traitement
WO2007126857A1 (fr) * 2006-03-27 2007-11-08 Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Inhibiteurs diamidine de tdp1

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997024367A1 (fr) * 1995-12-29 1997-07-10 Glaxo Group Limited Derives de 21-(2-oxo-tetrahydrofurane)-thiopregnane, leur procede de preparation et compositions pharmaceutiques les contenant
WO2001072293A2 (fr) * 2000-03-24 2001-10-04 Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Antiprogestins a activite partielle d'agonistes
WO2003014139A1 (fr) * 2001-08-08 2003-02-20 San Diego State University Foundation Compositions therapeutiques et procedes de criblage lies a la 3'-tyrosyl-adn-phosphodiesterase
WO2003015757A1 (fr) * 2001-08-16 2003-02-27 The Trustees Of The University Of Pennsylvania Synthese et utilisation de reactifs pour ameliorer la lipofection d'adn et/ou les therapies par medicaments et promedicaments a liberation lente
WO2006015369A2 (fr) * 2004-08-03 2006-02-09 The Johns Hopkins University Dosage d'identification d'inhibiteurs de topoisomerase
WO2006099095A2 (fr) * 2005-03-11 2006-09-21 The Government Of The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Inhibiteurs de ribosome utilises en tant qu'inhibiteurs de la tyrosyl-dna-phosphodiesterase
WO2007112121A2 (fr) * 2006-03-27 2007-10-04 Government Of The United States Of Americas, As Represented By The Secretary Composés à base de tétracyline et procédés de traitement
WO2007126857A1 (fr) * 2006-03-27 2007-11-08 Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Inhibiteurs diamidine de tdp1

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CARROLL F I ET AL: "STEROID DERIVATIVES OF PURINE-6 1H-THIONE 1A-C ANTI NEOPLASTIC MOUSE", JOURNAL OF HETEROCYCLIC CHEMISTRY, HETEROCORPORATION. PROVO, vol. 5, no. 6, 1 January 1968 (1968-01-01), pages 805 - 811, XP009104673, ISSN: 0022-152X *
DAVIES D R ET AL: "Insights into Substrate Binding and Catalytic Mechanism of Human Tyrosyl-DNA Phosphodiesterase (Tdp1) from Vanadate and Tungstate-inhibited Structures", JOURNAL OF MOLECULAR BIOLOGY, LONDON, GB, vol. 324, no. 5, 13 December 2002 (2002-12-13), pages 917 - 932, XP004457515, ISSN: 0022-2836 *
LI CHENGLONG ET AL: "Virtual screening of human 5-aminoimidazole-4-carboxamide ribonucleotide transformylase against the NCI diversity set by use of AutoDock to identify novel nonfolate inhibitors.", JOURNAL OF MEDICINAL CHEMISTRY 30 DEC 2004, vol. 47, no. 27, 30 December 2004 (2004-12-30), pages 6681 - 6690, XP002493223, ISSN: 0022-2623 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8716295B2 (en) 2010-10-27 2014-05-06 Yves Pommier Fluoroquinolone derivatives or sulfonamide moiety-containing compounds as inhibitors of tyrosyl-dnaphosphodiesterase (TDP1)
US11691963B2 (en) 2020-05-06 2023-07-04 Ajax Therapeutics, Inc. 6-heteroaryloxy benzimidazoles and azabenzimidazoles as JAK2 inhibitors
US11970494B2 (en) 2021-11-09 2024-04-30 Ajax Therapeutics, Inc. 6-heteroaryloxy benzimidazoles and azabenzimidazoles as JAK2 inhibitors

Similar Documents

Publication Publication Date Title
Rais et al. Discovery of 6-diazo-5-oxo-l-norleucine (DON) prodrugs with enhanced CSF delivery in monkeys: a potential treatment for glioblastoma
US8865686B2 (en) Tetracycline compounds as tyrosyl-DNA phosphodiesterase I inhibitors
JP5688288B2 (ja) がんの処置のための相乗的な医薬の組合せ
CN103119047B (zh) 三环类蛋白酶体活性增强化合物
US20110110886A1 (en) Small molecule inhibitors of autotaxin and methods of use
CN103402993A (zh) 用于疾病治疗的4，9-二羟基-萘并[2,3-b] 呋喃新型酯
JP2005533810A (ja) 新規な抗コレステロール組成物及びその使用方法
CA2829755A1 (fr) Traitement des troubles de la proliferation
CN101969954A (zh) 三芳基甲烷类似物及其在治疗癌症中的用途
US8716295B2 (en) Fluoroquinolone derivatives or sulfonamide moiety-containing compounds as inhibitors of tyrosyl-dnaphosphodiesterase (TDP1)
WO2004024140A1 (fr) Inhibiteurs de pompes d'efflux
CA2865983C (fr) Activite anticancereuse puissante par activation d'un compose double
CN114025766A (zh) 用于抑制gapdh的噁噻嗪化合物
WO2008124145A1 (fr) Composés stéroïdes, compositions et procédés de traitement
US20090298934A1 (en) Diamidine Inhibitors of TDP1
US20070213312A1 (en) Hif-1 Inhibitors and Methods of Use Thereof
JPH06510280A (ja) タンパクキナーゼｃ阻害および新規化合物バラノール
CN105189469A (zh) 2-氨基-3,4-二氢喹唑啉衍生物及其作为组织蛋白酶d抑制剂的用途
Zhang et al. The activity of novel BCR-ABL small-molecule degraders containing pyrimidine rings and their role in overcoming drug resistance
WO2012129057A2 (fr) Inhibiteurs de ssh-2 (slingshot-2) et procédés pour les préparer et les utiliser
CN102875479B (zh) 喹唑啉二酮衍生物及其制备方法、其药物组合物及用途
Supuran et al. Carbonic anhydrase II as target for drug design
Ma et al. ML162 derivatives incorporating a naphthoquinone unit as ferroptosis/apoptosis inducers: Design, synthesis, anti-cancer activity, and drug-resistance reversal evaluation
Zhu et al. Synthesis and bioevaluation of novel stilbene-based derivatives as tubulin/HDAC dual-target inhibitors with potent antitumor activities in vitro and in vivo
US20240109852A1 (en) Ferroptosis-HDAC Inhibitor Hybrid Anticancer Agents

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08742655

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08742655

Country of ref document: EP

Kind code of ref document: A1