CN1795187A - Novel ido inhibitors and methods of use - Google Patents

Abstract

Compounds, compositions and methods for the treatment of malignancy are disclosed.

Description

Novel IDO inhibitor and using method thereof

The application enjoys U.S. Provisional Application number 60/527 according to 35U.S.C. § 119 (e) request, 449 (proposing) U.S. Provisional Application number 60/458 on December 5th, 2003, the right of priority of 162 (proposing on March 27th, 2003), the full content of two pieces of applications is here introduced.

Invention field

The present invention relates to oncology.In particular, the present invention provides the using method of novel chemotherapeutics and this medicament for tumor treatment.

Background of invention

That tumour is peculiarly expressed is atypical, have potential immunoreactive antigen, jointly is called as tumour antigen.More and more evidences shows, immunity system can not be set up effective response to the tumour that growth day by day increase and can not be attributed to and lack the tumour immunity antigen of having understood.The inhibition immune response of tumour is understood seldom, tumour is escaped the never further investigation of mechanism of immunological surveillance.Recently, evidence suggests that cytotoxin T cell is by by indoleamine 2, the reduction of the tryptophane partial concn that the activity of 3-dioxygenase (IDO) causes and become tolerance.

IDO be a kind of in the katabolism of tryptophane the rate-limiting step in the catalysis tryptophane katabolism.This kind of enzyme is structurally completely different with the catabolic tryptophane dioxygenase of tryptophane (TDO) of being responsible for diet in liver.IDO is a kind of IFN-γ target gene, and thinks and understood certain effect [Mellor and Munn, (1999) current immunology (Immunol.Today), 20:469-473] in immunomodulatory.The active increase of IDO has reduced the tryptophan levels of local cells environment.IDO inducing in antigen presenting cell (in this cell IDO by IFN-γ regulate) blocked the activation of T cell, and be particularly responsive to the loss of tryptophane.The T cell must just can become activated through the cell fission of 1-2 wheel, and still for the loss of corresponding tryptophane, they are trapped in the G1.Like this, suggestion suppresses to promote the T of cytotoxin T cell development with IDO _H1 reaction.

The material evidence of the effect of IDO in immunosuppression is by 1-methyl-tryptophane (1MT)--a kind of special, bioactive IDO inhibitor [Cady and Sono, (1991) biological chemistry and biophysics archives (Arch.Biochem.Biophys.), 291:326-333] performance prove, elicit allogeneic mouse embryo's MHC restriction and the cell-mediated rejection effect [Mellor etc. of T, (2001) natural immunity (Nat.Immunol.), 2:64-68; Munn etc., (1998) science (Science.), 281:1191-93].The high level that IDO in this effect and the placental trophoblasts expresses is consistent [Sedlmayr etc., (2002) molecular human breedings (Mol.Hum.Reprod.), 8:385-391].

It should be noted that the IDO activity is trend [Yasui etc., (1986) national science research institute journal (Proc.Natl.Acad.Sci.), the U.S., the 83:6622-26 that improves constantly in human tumor or cancer patients's body; Taylor and Feng, (1991) FASEB are J.5:2516-22].Since IDO can regulate immune response, will draw the connotation of a reasonable logic so: the increase of IDO in tumour may promote immunosuppression reaction [Mellor and the Munn of tumour, (1999) current immunology (Immunol.Today), 20:469-473; Munn etc., (1999) experimental medicine periodical (J.Exp.Med.), 189:1363-72; Munn etc., (1998) science (Science.), 281:1191-93].This possibility shows many tumours of its feature by the reduction of the favourable immunologic function that can limit its malignant development, comprises that the observations of mammary cancer is supported.For example, promote the T that cytotoxin T cell produces _H1 reaction (its sign is to produce IFN-γ) is suppressed during cancer worsens.From the hypothesis that these data drew being exactly, if IDO impels cancer to worsen by the activation that weakens the T cell, is the immunosuppression reaction of media by changing with IDO so, and IDO restraining effect in animal body should reduce growth of tumor.But in mouse model, the transmission of IDO inhibitor 1-methyl-tryptophane (1MT) is just slowed down and can not be stoped growth of tumor [Friberg etc., (2002) international cancer periodical (Int.J.Cancer), 101:151-155; United States Patent (USP) (US Patent), 6,482,416].

Cell signal conversion promptly a series ofly comes from extracellular effect and causes into consequence in the cell, is an aspect of cell function under standard state and the sick state.The protein that measures the signal transducers effect greatly is identified, comprises that acceptor and nonreceptor tyrosine kinase, Phosphoric acid esterase and other have enzyme or regulate active molecule.These molecules prove that usually it has certain capabilities to form a kind of signal mixture that can change hyperplasia with other protein bound.

The anomalous signals transduction may cause conversion, the advolution of malignant tumour.Therefore, can use the inhibitor in signal transduction path to treat cancer.In the past few years, develop many signal transduction inhibitors (STIs), nowadays studied the ability that they suppress tumor growth.

Summary of the invention

According to an aspect of the present invention, provide new indole amine 2, the activity of 3-dioxygenase (IDO) inhibitor.Novel cpd is by selecting in molecular formula (I) and the molecular formula (II):

Molecular formula (I): Wherein, R ₁Be H or low alkyl group; R ₂Be H; R ₃Be from following group, to select: (a) R wherein _AAnd R _BFrom group H and alkyl, select independently; (b)

R wherein _CFrom group H and alkyl, select; (c)

Wherein n is the integer of 1-10, R _DBe molecular formula: The carboline substituting group; And (f)

R wherein _AAnd R _BFrom group H and alkyl, select independently; Perhaps R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group:

With The part of the ring of selecting, R in the base _EBe alkyl or alkyl-Q, Q represents molecular formula:

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Upgrading becomes beta-carboline derivatives when combining representative (i), works as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also can be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin; Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

According to a further aspect in the invention, provide a kind of in patient's interior therapeutic method for cancer.This method comprise use significant quantity can accept to contain in the mounting medium at least a indoleamine 2 in pharmacy, the pharmaceutical composition of 3-dioxygenase (IDO) inhibitor, preferably a kind of novel inhibitors of the present invention.

In another situation of the present invention, the method that the patient that being provided at needs it carries out cancer therapy on one's body.This method comprises it being to allow the patient use, side by side or according to priority, and at least a indoleamine 2 of significant quantity, 3-dioxygenase (IDO) inhibitor and at least a signal transduction inhibitor (STI).In a particular embodiment of the present invention, at least a STI selects from bcr/abl kinase inhibitor, Urogastron (EGF) acceptor inhibitor, her-2/neu acceptor inhibitor and farnesyl tranfering enzyme inhibitor (FTIs).Compound can the pharmaceutically acceptable carrier medium in administration.

The present invention also has an embodiment to be exactly, another method that the patient that being provided at needs carries out cancer therapy on one's body.This method comprises to the patient to be used, side by side or according to priority, and at least a indoleamine 2 of significant quantity, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutics.In a specific embodiment of the present invention, above-mentioned at least a chemotherapeutics is selected all kinds of from following: taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.This compound can carry out in the pharmaceutical acceptable carrier medium.

Again according to an aspect of the present invention, a kind of method is provided, patient at needs carries out cancer therapy on one's body, wherein by using to the patient, side by side or according to priority, significant quantity at least a except that a kind of IDO inhibitor immunomodulator and at least a cytotoxin chemotherapeutics or at least a STI of significant quantity.In an embodiment, at least a immunomodulator is selected all kinds of from following: CD40L, B7, B7RP1, ant-CD40, anti-CD38, anti-ICOS, 4-IBB part, dentritic cell Theratope, IL2, IL12, ELC/CCL19, SLC/CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFNa/b, M-CSF, IL-3, GM-CSF, IL-13 and anti-IL-10.In another kind of embodiment, above-mentioned at least a cytotoxin chemotherapeutic is selected all kinds of from following: taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.

The present invention also has a kind of situation to be exactly, and a kind of method is provided, and carries out the treatment of chronic viral infection on one's body its patient of needs, by using to the patient, side by side or according to priority, at least a indoleamine 2 of significant quantity, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutics.Above-mentioned at least a chemotherapeutics can be selected all kinds of from following: taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.Active agents adopts or does not adopt the administration of pharmaceutically acceptable carrier medium.Slow virus sexuality arbitrarily below method of the present invention can be used for the treatment of: hepatitis C virus (HCV), Human papilloma virus HPV (HPV), cytomegalovirus (CMV), epstein-Barr virus (EBV), varicella zoster virus, Coxsackie virus and human immunodeficiency virus (HIV).

The concise and to the point description of illustration

Fig. 1 provides the synthetic method of thiohydantoin derivative among the present invention.

Fig. 2 provides indoles deutero-route on C-4 and N-1 position among the present invention.

Fig. 3 provides the synthetic route of the indoles that C-6 replaces among the present invention.

Fig. 4 provides the synthetic route of C-4 among the present invention, C-5 and the trisubstituted indoles of C-6.

Fig. 5 provides the optionally synthetic route of thiohydantoin derivative among the present invention.

Fig. 6 provides the synthetic route of brassinin derivative among the present invention.

Fig. 7 provides the synthetic route of the beta-carboline derivatives that C-5 and C-6 replace among the present invention.

Fig. 8 provides the synthetic route of the beta-carboline derivatives that the C-3 alkyl replaces among the present invention.

Fig. 9 provides the optionally synthetic route of beta-carboline derivatives among the present invention.

Figure 10 provides the synthetic route of 3-amino among the present invention-6/7-bromo-2-naphthoic acid.

Figure 11 provides the synthetic route of C-6 among the present invention, C7 and the trisubstituted 3-amino of C-8-2-naphthoic acid.

Figure 12 provides the optionally synthetic route of C-6 among the present invention, C-7 and the trisubstituted 3-amino of C-8-2-naphthoic acid.

Figure 13 provides the synthetic route of brassilexin derivative among the present invention.

Figure 14 provides the optionally synthetic route of the brassilexin derivative that replaces among the present invention.

Figure 15 provides the synthetic route of the brassilexin derivative that N-replaces among the present invention.

Figure 16 provides the synthetic route of cyclopropyl tryptophane among the present invention.

Figure 17 provides the synthetic route of '-aziridino tryptophan derivative among the present invention.

Figure 18 provide mooring among the present invention competitiveness usefulness/synthetic route of noncompetitive derivative.

Figure 19 A and 19B are the graphic representation of the enzyme experimental result of IDO inhibitor.A) 1MT and B) under the ever-increasing situation of concentration of MTH-Trp, obtain the spherical nonlinear regression analysis of the enzyme dynamic date of relevant human body IDO.Utilize Prism4 software package (GraphPad) to draw and analyze.The fit value of the Ki of 1MT be 34.6 μ M MTH-Trp then be 11.4 μ M.

Figure 20 is relevant 1MT to IDO, 1MT to TDO and MTH-Trp to the IDO inhibitor result of experiment graphic representation on the cell base of the 2log dose escalation study of IDO.Utilize Prism4 data analysis system (GraphPad) to draw, determine the gradient and EC50 value by nonlinear regression analysis.

Figure 21 is the synoptic diagram in kynurenine metabolism path.IDO is a kind of oxydo-reductase that liver is outer, interferon-brings out that comes from.IDO reaction product---the N-formylkynurenine promptly is hydrolyzed into kynurenine.Kynurenine is distributed between the space of tissue and blood, because enzyme has only very little or do not have activity to make kynurenine further metabolic in tissue.The main path of removing kynurenine is to discharge with urine change into xanthurenic acid in liver and/or kidney after, although it also is the media [Takikawa etc. that produce nicotine adenine dinucleotide (NAD) in biosynthetic pathway, (1986) biological chemistry periodical (J.Biol.Chem.), 261:3648-3653; Thomas and Shi Tuokeer, (1999) redoxomorphism report (Redox.Report), 4:199-220].

Figure 22 A-D is the color atlas of mice serum HPLC analytical results.Serum is cultivated the blood sample of gathering and is removed blood clot and is prepared from by spending the night under 4 ℃.Remove deproteinize by the TCA precipitating action.By 20%MeOH, 5% acetonitrile, 10mMKPO ₄(pH5.3), 0.15mMEDTA constitute etc. the 250mm * 4.5mm silver 5 μ C18 posts (Phenomenex) in the degree snubber go up sample dissolution.Serum sample prepares in the FVB class mouse body for the treatment of by following each step: A) not treatment (being incorporated in the serum with every kind of control compound below the 30 μ M: tryptophane, kynurenine and 1MT); B) not treatment (can detect 55 μ M serum tryptophanes); C) 24 hours (can detect and reach inducing of 5.6 μ M kynurenines) of LPS test; D) the subcutaneous injection of 1MT piller 3 days (can detect the 1MT of 104 μ M).When 6.66 minutes and 14.5 minutes, regulate output sensitivity (Y-axis), and the peak height of each expection of optimization.Ky is a kynurenine, and W is a tryptophane, and 1MT is 1-methyl-tryptophane.

Figure 23 illustrates the graphic representation that the merging of the gross tumor volume of MMTVneu mouse changes, and this mouse simulated treatment (treating) or with 1MT, L-744,832 (FTI), 1MT and L-744,832 and the chemotherapeutics that 1MT arranged or do not have a 1MT treated.Each data point is determined by single mouse that all column is represented the mean number of data point, be listed in graphic representation below.

Figure 24 is the result curve figure that the external biological chemistry is useful for examination IDO inhibitor material standed for person.Relevant kynurenine quantity data under the situation that inhibitor exists is provided.

Figure 25 A and 25B are the result curve figure that is useful for examination IDO inhibitor material standed on the cell base.Among Figure 25 A, provide relevant kynurenine quantity data under the non-existent situation of inhibitor.Among Figure 25 B, provide the data of relevant fluorescence aspect, the generation (that is IDO activity) of expression kynurenine.Cell comes transfection with empty expression vehicle (vehicle) or with the expression vehicle of the cDNA that contains IDO.

Figure 26 provides the graphic representation of the thiohydantoin derivative of the indoles amine that the experiment on the cell base is used for examination IDO inhibitor material standed for.Cell comes transfection with empty expression vehicle (vehicle) or with the expression vehicle that contains IDO or TDO.In order to contrast, to express the vehicle cells transfected with IDO and also will under the situation that 1MT exists, chemically examine.

Figure 27 is the chart of several IDO inhibitor, comprises suppressing IDO and the active ability of TDO when concentration is 250 μ M in its structure and their experiments on cell base.

Figure 28 offer curves figure confirms the toxicity of the IDO inhibitor of several newborn mammary gland (top board) that shift or prostate gland (base plate) cancer cells.Cell is not treated (Untx) or is treated with the inhibitor of 100 μ M.

Figure 29 is the graphic representation of total variation of the gross tumor volume of explanation MMTVneu mouse, and this mouse simulated treatment (treating) or treated with 1MT, taxol (taxol ), 1MT and taxol (taxol ), Platinol or 1MT and Platinol.Each data point is determined by single mouse that all column is represented the mean number of data point, be listed in graphic representation below.

The present invention elaborates

In an embodiment of the invention, provide many demonstration IDO to suppress active new compound. In addition, the pharmaceutical composition that is made of this kind compound and the using method that suppresses the tumour growth thereof are included among the present invention.

Another embodiment of the invention is, a kind of combined therapy scheme of using signal transduction inhibitor (STI) and IDO inhibitor that comprises is provided, and it is effective to the growth that suppresses tumour.

And according to a further aspect in the invention, provide another kind to comprise to use the combined therapy scheme of chemotherapeutics and IDO inhibitor. It also is effective to the growth that suppresses tumour.

The present invention also has in the concrete enforcement mode, provides a kind of combined therapy scheme to be used for the treatment of chronic virus and has infected, and this scheme comprises uses IDO inhibitor and chemotherapeutics.

I. definition

Term " IDO inhibitor " refers to suppress indoleamine 2,3-add dioxygenase (IDO) thus activity reverse a kind of medicine with the Immunosuppression reaction of IDO mediation. The IDO inhibitor may be competitive, and is non-competing property or irreversible. " competitive IDO inhibitor " is a kind of compound, can reversibly suppress in catalytic site the activity (such as but not limited to, 1-methyl-tryptophan) of IDO enzyme; " non-competing property IDO inhibitor " is a kind of compound that can reversibly suppress at non-catalysis position the IDO enzymatic activity (such as but not limited to, two nitrogen fluorenes); Also have, " irreversible IDO inhibitor " is a kind of compound that irreversibly destroys the IDO enzymatic activity by forming covalent bond with enzyme (such as but not limited to, cyclopropyl/'-aziridino tryptophan derivative).

IDO inhibitor among the present invention may include but not limited to: (known) IDO inhibitor of i) having determined before, such as but not limited to: 1MT (1MT; Sigma-Aldrich; The St. Louis, the Missouri State), β-(3-benzo furyl)-DL-propylamine acid (Sigma-Aldrich), β-(3-benzo (b) thiophene base)-DL-propylamine acid (Sigma-Aldrich), 6-nitro-L-Trp (Sigma-Aldrich), indoles 3-methyl alcohol (LKT laboratory, Sao Paulo, the Minnesota State), 3,3'Diindolylmethane (LKT laboratory), epi-nutgall catechu acid Gallate acid salt (LKT laboratory), 5-bromo-4-chloro-indoxyl 1, the two acetic acid esters (Sigma-Aldrich) of 3-, 9-vinyl carbazole (Sigma-Aldrich), A Ximeixin (Sigma-Aldrich), 5-bromine generation-DL-Trp (Sigma-Aldrich), the 5-bromine is for the two acetic acid salt (Sigma-Aldrich) of indoxyl; And ii) the new IDO inhibitor among the present invention. The present invention's preferred embodiment is that this IDO inhibitor comprises the new IDO inhibitor of invention.

" signal transduction inhibitor " is a kind ofly can suppress one or more important step in the signal path selectively in the normal function of cancer cells, thereby causes apoptotic medicine.

Signal transduction inhibitor among the present invention (STIs) includes but not limited to (i) bcr/abl kinase inhibitor, for example, and STI 571 (Gleevec); (ii) Urogastron (EGF) acceptor inhibitor, for example, kinase inhibitor (Iressa, SSI-774) and antibody (Imclone:C225[Goldstein etc., (1995), Clinical Cancer Research (Clin CancerRes.), 1:1311-1318], and Abgenix:ABX-EGF); (iii) her-2/neu acceptor inhibitor, for example, Trastuzumab ^TM(Herceptin), and farnesyl tranfering enzyme inhibitor (FTI) is as L-744,832[Kohl etc., (1995), country's medicine (Nat Med.), 1 (8): 792-797]; The (iv) inhibitor in the kinases of Akt series or Akt path, for example, rapamycin is (as (2000) cancer research (Cancer Res.) referring to works such as Sekulic, 60:3504-3513); (v) cell cycle kinase inhibitors, for example, flavopiridol and UCN-01 are (as the carcinostatic agent (Curr.Med.Chem.Anti-Canc Agents) of (2003) current medical chemistry of writing referring to Sausville, 3:47-56); (vi) phosphatidyl inositol kinase inhibitor, for example, LY294002 is (as (1994) the biological chemistry periodical (J.Biol.Chem.) referring to books such as Vlahos, 269:5241-5248).

" the treatment significant quantity " of compound or pharmaceutical composition is meant a kind of dose, can regulate tumor growth or transfer in animal body, particularly in human body, include but not limited to reduce tumor growth or size or give the formation of the animal tumor growth in vivo that forms without any tumour before the anti-administration, promptly preventative taking.

" pharmacy is acceptable " is meant through the approval of the administration that lists in federation or national government or American Pharmacopeia or other pharmacopeia of having generally acknowledged usually, is used for animal especially for the mankind.

" carrier " is meant, for example, thinner, assistant agent, vehicle, auxiliary agent or vehicle utilize their promoting agents of the present invention to be applied.This pharmaceutical carriers can be a sterilising liq, as water or oil, comprises the oil in oil, animal, vegetables or synthetic source, as peanut oil, soybean oil, mineral oil, sesame oil or the like.Water or salt brine solution and D/W and glycerine solution all are the relatively more suitable carriers that is used as, in particular for Injectable solution.The carrier that makes up a prescription that some are suitable is set forth (" Remington ' s Pharmaceutical Sciences " by E.W.Martin.) to some extent in " the pharmacy science of Lei Mingdun " that E.W. Martin is write.

" side by side " meaning is to be simultaneously on (1) time, or (2) different time in same treatment plan process.

" according to priority " be meant that a kind of composition of using this method then uses another kind of composition.Use after a kind of composition, next composition can be used after first kind of composition basically at once, and perhaps next composition can be through using after one period effective time cycle after first kind of composition again; This effective time cycle is meant from using of first kind of composition and obtains the time of effect to greatest extent.

" alkyl " is meant not the hydrocarbon polymer part that replaces or replace, and it contains has an appointment 1 to about 10 carbon atoms or contain and have an appointment 1 to about 25 carbon atoms, and these carbon atoms can be straight chain, side chain or cyclic alkyl.When being substituted, alkyl may be substituted on any available tie point.Representational not substituted radical comprises alkyl, as methyl, ethyl, propyl group, sec.-propyl, n-butyl, t-butyl, isobutyl-, amyl group, hexyl, isohexyl, heptyl, 4,4-dimethyl amyl group, octyl group, 2,2,4-tri-methyl-amyl, nonyl, decyl, undecyl, dodecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, tetracosyl, pentacosyl or the like; Aryl is as phenyl, tolyl, xylyl, naphthyl, biphenyl, tetraphenyl or the like; Aralkyl is as phenmethyl, styroyl, hydrocinnamyl, benzene butyl, benzene hexyl, naphthalene octyl group or the like; And cycloalkyl, as cyclopropyl, cyclobutyl, cyclohexyl, suberyl, ring octyl group or the like.Representational substituting group may include but not limited to following one or a following groups: and halogeno-group (as F, Cl, Br, I), alkylhalide group is (as CCl ₃Or CF ₃), alkoxyl group, alkylthio, hydroxyl, carboxyl (COOH), carbonyl (C (=O)), epoxy group(ing), alkoxy carbonyl (C (=O)-OR), the alkyl-carbonyl oxygen base (OC (=O)-R), amino (NH ₂), carbamyl (NH ₂C (=O)-or NHRC (=O)-), urea groups (NHCONH ₂), the alkane urea groups (NHCONHR) or mercaptan (SH), the R in the wherein above-mentioned substituting group represents a hydrocarbyl portion.Pressing here, defined hydrocarbyl group (part) also may comprise one or more carbon-carbon double bond or one or more carbon carbon triple bond (that is, hydrocarbyl group may be undersaturated).Representational unsaturated alkyl group comprises alkenyl, as allyl group and vinyl.Hydrocarbyl group also may be interrupted by at least one oxygen, nitrogen or sulphur atom.

Term " halogen ", " halo " reach " halogenide " and are meant chlorine, bromine, fluorine or iodine.

" low alkyl group " is meant the alkyl that contains 1 to 4 carbon atom of having an appointment, as methyl, ethyl, propyl group, butyl and sec.-propyl.In a specific implementations, low alkyl group is meant methyl.

II. have IDO and suppress active novel cpd and using method thereof

According to saying of the present invention, provide novel cpd, thereby it can suppress the active tumor growth that suppresses of IDO with following molecular formula:

Molecular formula (I):

R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃From following group, select: (a)

R wherein _AAnd R _BFrom H and hydrocarbyl group, select independently; (b)

R wherein _CFrom H and hydrocarbyl group, select; (c) Wherein n is from 1 to 10 integer, R _DBe molecular formula: The carboline substituting group; (f) R wherein _AAnd R _BFrom H and hydrocarbyl group, select respectively; Perhaps R ₂And R ₃Combine and represent molecular formula (I) and pyrroles's part condensed and from group:

With The part of the middle ring of selecting, wherein R _EBe an alkyl or alkyl-Q, Q represents molecular formula: Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become N-replacement brassilexin derivative when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin; And molecular formula (II):

Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

An embodiment of the invention are that the novel cpd of molecular formula (I) may further show its feature by following supplementary condition: 1) if R ₃Be substituting group (a) and R _ANot H, R so _BBe alkyl, perhaps at least one of X, Y or Z is not H, NO ₂, COH, CH ₂OH or CH ₃2) if R ₃Be substituting group (a) and R _BBe H, R so _ABe H, perhaps X, Y or Z's is not H, NO one of at least ₂, COH, CH ₂OH or CH ₃3) if R ₃Be that substituting group (a) and X, Y and Z are from H, NO ₂, COH, CH ₂OH or CH ₃Select in the group, so R _ABe H, perhaps R _BIt is alkyl.

And another embodiment of the invention is, the novel cpd of molecular formula (I) may further show its feature by following supplementary condition: work as R ₃When being (f), Y is a halogen.

A specific embodiment of the present invention is to work as R ₃When being (f), X and Z are H, and Y is Br, R ₁Be-CH ₃, R ₂Be H, R _ABe-CH ₃Or-CH ₂-CH=CH ₂, R _BBe H.

The compound of molecular formula (I), wherein R ₃Selection from the group (a) and (b) and (f), and molecular formula (II) all think can competitive inhibition IDO activity, and the compound of molecular formula (I), wherein R ₃From group (c), (d) with select (e), perhaps R ₂And R ₃As if in conjunction with forming group (i), one of (ii) and (iii), noncompetitive suppresses the activity of IDO.

The present invention also is provided to contain in the pharmaceutical acceptable carrier to have from molecular formula (I) and the pharmaceutical composition of at least one IDO inhibitor of a molecular formula of (II) selecting.This pharmaceutical composition can use for the patient who needs it to treat cancer with certain treatment significant quantity.

In addition, the present invention also provides a kind of treatment method for cancer, uses the IDO inhibitor of at least a molecular formula that contains from molecular formula (I) and select (II) of certain treatment significant quantity promptly for the patient who needs it.

Can use the scheme among the present invention to treat the cancer that includes but not limited to following organ-tissue: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), an endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

III. the conjoint therapy that is used for cancer therapy

The present invention also provides the inhibition method of tumour.According to the present invention, find that the combination of IDO inhibitor and signal transduction inhibitor (STI) can produce synergy and suppress growth of tumor.Therefore, the invention provides a kind of pharmaceutical composition and treat the cancer patients, said composition contains at least a IDO inhibitor and at least a STI in pharmaceutical acceptable carrier.A kind of cancer patients's of treatment method also is provided in addition, promptly uses at least a IDO inhibitor of significant quantity and linked together with at least a STI.Suitable IDO inhibitor comprises the compound of a molecular formula that contains from molecular formula (I) and select (II) and demonstrates any compound of IDO inhibitory activity.Adaptive STI as above writing, includes but not limited to: (i) bcr/abl kinase inhibitor, for example, STI 571 (Gleevec); (ii) Urogastron (EGF) acceptor inhibitor, for example, kinase inhibitor (Iressa, SSI-774) and antibody (Imclone:C225[Goldstein etc. (1995), Clinical Cancer Research (Clin CancerRes.), 1:1311-1318], and Abgenix:ABX-EGF); (iii) her-2/neu acceptor inhibitor, for example, Trastuzumab ^TM(Herceptin), and farnesyl tranfering enzyme inhibitor (FTI), L-744 for example, 832[Kohl etc., (1995), country's medicine (Nat Med.), 1 (8): 792-797]; The (iv) kinase inhibitor of Akt series or Akt path, for example, rapamycin is (as (2000) cancer research (Cancer Res.) referring to works such as Sekulic, 60:3504-3513); (v) cell cycle kinase inhibitors, for example, flavopiridol and UCN-01 are (as the carcinostatic agent (Curr.Med.Chem.Anti-Canc Agents) of (2003) current medical chemistry of writing referring to Sausville, 3:47-56); (vi) phosphatidyl inositol kinase inhibitor, for example, LY294002 is (as (1994) the biological chemistry periodical (J.Biol.Chem.) referring to books such as Vlahos, 269:5241-5248).

An embodiment of the present invention is, at least a IDO inhibitor and at least a STI can be side by side or be administered to the patient according to priority.In other words, can use at least a IDO inhibitor earlier and use at least a STI again; Perhaps at least a IDO inhibitor and at least a STI can use simultaneously.In addition, when IDO inhibitor that uses more than one and/or STI, can use these compounds by any order.

The cancer that can use the scheme for combining among the present invention to treat includes but not limited to the cancer of following organ-tissue: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), between endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

Except IDO, recognize that also other molecules also have immunoregulatory effect.These molecules also may be the potential targets that is used for suppressing tumor growth among the cancer patients.Thereby the present invention also provides treatment cancer patients's integrated processes, except using IDO inhibitor and at least a chemotherapeutics, uses at least a immunomodulator again.Adaptive can include but not limited to by immunomodulator used in this invention: costimulatory molecules, and as CD40L, B7 and B7RP1; Be total to the activation monoclonal antibody (mAbs) of costimulatory receptor, as ant-CD40, anti-CD38, anti-ICOS and 4-IBB part; Dentritic cell antigen load (external or body in); The dentritic cell cancer vaccine; Cytokine/chemokine is as IL1, IL2, IL12, IL18, ELC/CCL19, SLC/CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFNa/b, M-CSF, IL-3, GM-CSF, IL-13 and anti-IL-10; Bacteria lipopolysaccharide (LPS); And the immunostimulation oligonucleotide (for example, CpG DNA polymkeric substance (referring to, as Verthelyi and Zeuner (2003), treatment immunology (Tr.Immunol.), 24:519-522)).Adaptive chemotherapeutics includes but not limited to cytotoxicity chemotherapeutics and signal transduction inhibitor (STIs).

According to the present invention, find that also the combination of IDO inhibitor and chemotherapeutics can act synergistically and suppress tumor growth.Therefore, the invention provides a kind of pharmaceutical composition and treat the cancer patients, said composition contains at least a IDO inhibitor and at least a chemotherapeutics in pharmaceutical acceptable carrier.A kind of cancer patients's of treatment method also is provided in addition, promptly uses at least a IDO inhibitor of significant quantity and combine with at least a chemotherapeutics.Adaptive IDO inhibitor comprises the compound of a molecular formula that contains from molecular formula (I) and select (II) and demonstrates any compound of IDO inhibitory activity.Adaptive chemotherapeutics, as above writing, include but not limited to: taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.

A specific embodiment of the present invention is, at least a IDO inhibitor and at least a chemotherapeutic can be side by side or be administered to the patient according to priority.In other words, can use at least a IDO inhibitor earlier and use at least a chemotherapeutic again; Perhaps at least a IDO inhibitor and at least a chemotherapeutic can be used simultaneously.In addition, when IDO inhibitor that uses more than one and/or chemotherapeutic, can use these compounds by any order.

Can use the assembled scheme among the present invention to treat the cancer that includes but not limited to following organ-tissue: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), an endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

IV. treat the conjoint therapy of chronic viral infection

In addition, the invention provides the chronic viral infection that a kind of pharmaceutical composition is used for the treatment of the patient, it contains at least a IDO inhibitor, at least a cancer treatment drugs and dispensable at least a antiviral agent in pharmaceutical acceptable carrier.In addition, provide a kind of method for the treatment of patient's chronic viral infection by the composition of using above-mentioned significant quantity.

Suitable IDO inhibitor comprises the compound of a molecular formula that contains from molecular formula (I) and select (II) and demonstrates any compound of IDO inhibitory activity.

Suitable chemotherapeutic is any compound that demonstrates antitumour activity, includes but not limited to: alkylating agent (for example, mustargen such as Chlorambucil, endoxan, isofamide, dichloromethyldiethylamine, melphalan and uracil mustard; Aziridine such as tespamin; Methane sulfonate class such as busulfan; Nitrosourea such as Carmustine, lomustine and U-9889; Platinum complex such as Platinol and NSC-241240; The alkide of biological reducing such as mitomycin, procarbazine, dacarbazine and altretamine); DNA splitting of chain agent (for example, bleomycin); Topoisomerase II inhibitor (for example, amsacrine, gengshengmeisu, daunomycin, jaundice element, mitoxantrone, adriamycin, etoposide and Vumon); The DNA minor groove binding (for example, plicamydin); Metabolic antagonist (for example, folic acid antagonist such as methotrexate and TMQ; Pyrimidine antagonistic such as Fluracil, fluorodeoxyuridine, CB3717, azacitidine, cytosine arabinoside and floxuridine; Purine antagonistic such as purinethol, 6-Tioguanine, fluorine draw and reach shore, pentoside; Asparaginase; And ribonucleotide reductase such as hydroxyurea); Tubulin interaction agent (for example, vincristine(VCR), vinealeucoblastine(VLB) and taxol (taxol)); Hormone drug (for example, oestrogenic hormon; Conjugated estrogen; Ethinylestradiol; Stilboestrol; Chlortrianisen; Idenestrol; Progesterone such as Hydroxyprogesterone caproate bp 98, Zytron and megestrol; Male sex hormone such as testosterone, testis pernicious influence propionic ester, fluoxymesterone and methyl testosterone); Adrenocortical steroid (for example, prednisone, dexamethasone, prednisolone and prednisolone base EP/Bp93 micro); Prolan B releasing agent or antagonists of gonadotropin-releasing hormone (for example, sharp special acetate of bright dried meat and goserelin acetate); Hormone antagonist antigen (for example, tamoxifen, antiandrogen such as flutamide; Antiadrenergic drug such as mitotane and amino are led the spirit of sleeping).Chemotherapeutics is preferably selected from following medicine: taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.

Suitable antiviral agent includes but not limited to: acycloguanosine; Gangcyclovir; Phosphorylcholine calcium; Virazole; And anti-retrovirus medicine such as nucleoside analog reverse transcriptase inhibitors are (for example, Azidothymidine (AZT), dd1, ddC, 3TC, d4T), non-nucleoside reverse transcriptase inhibitor (for example, efavirenz, nevirapine), nucleoside analog reverse transcriptase inhibitors and proteinase inhibitor.

A specific embodiment of the present invention is that at least a IDO inhibitor and at least a chemotherapeutics can simultaneously or be used to the patient in order.In other words, can use at least a IDO inhibitor earlier and use at least a chemotherapeutic again; Perhaps at least a IDO inhibitor and at least a STI can use simultaneously.In addition, when IDO inhibitor that uses more than one and/or chemotherapeutic, can use these compounds by any order.Similarly, antiviral drug also can be used at any time.

The compound of this combination therapy also can the drug treatment local infection.Especially, can use at least a IDO inhibitor, at least a chemotherapeutics, dispensable at least a antiviral drug and treat skin infections, as zoster and wart.This compound can be used in pharmaceutical acceptable carrier, and these carriers include but not limited to: gel, creme, lotion, ointment, powder, aerosol and traditional with medicinal application other forms to the skin.

Can use the chronic viral infection that conjoint therapy is treated among the present invention, include but not limited to the disease that following virus causes: hepatitis C virus (HCV), human body papillomavirus (HPV), cytomegalovirus (CMV), the single virus of bleb (HSV), epstein-Barr virus (EBV), the banded exanthema virus of varicella, Coxsackie virus, HIV (human immunodeficiency virus) (HIV).

Particularly, parasitiation (for example malaria) also can be treated with above-mentioned method, and wherein the compound of the treatment parasitiation of Gong Rening optionally adds, and replaces antiviral drug.

V. pharmaceutical composition and compound uses

Pharmaceutical composition among the present invention also can come administration by other suitable paths, for example, and injection, oral, lung, nasal cavity or other administering modes.Substantially, inter alia, pharmaceutical composition of the present invention comprises the acceptable diluent of pharmacy, sanitas, solubilizing agent, emulsifying agent, adjuvant and/or carrier.This composition may comprise the thinner (for example, Tris-HCl, acetate, phosphoric acid salt) of different buffer content things, pH and ionic strength; And additive such as sanitising agent and solubilizing agent (for example, tween 80, polysorbate 80), antioxidant (for example, xitix, sodium metabisulfite), sanitas (for example, thiophene mercury spreads, phenylcarbinol) and expanded material (for example, lactose, N.F,USP MANNITOL).Said composition can combine with the particle type preparation or the liposome of polymerizable compound such as poly(lactic acid), polyglycolic acid etc.This composition can influence the interior release rate of physical condition, stability, body and the interior removing speed of body of the component of Chinese traditional medicine composition of the present invention.Referring to thunderous bright " pharmacy science " (Remington ' sPharmaceutical Sciences), the 18th edition (1990, mark publishing company, Easton, Pennsylvania 18042) 1435-1712 page or leaf is incorporated herein by reference here.For example, pharmaceutical composition of the present invention can perhaps also can be prepared (as lyophilized powder) with the form of dry powder with the form preparation of liquid.

And another embodiment is, pharmaceutical composition of the present invention can transmit in controlled release system, such as utilizing vein to inject, and transplantable osmotic pump, transdermal patch, liposome or other are taken mode.Under special circumstances, can use pump (referring to Lange, on seeing; Sefton, CRC comment for reference bio-pharmaceuticals engineering (CRC Crit.Ref.Biomed.Eng.) (1987) 14:201; Buchwald etc., surgery (1980) 88:507; Saudek etc., New England Journal of Medicine (N.Engl.J.Med.) (1989) 321:574).Another kind of embodiment, can utilize polymeric material (referring to the medical applications (MedicalApplications of Controlled Release) of controlled release, Lange and Huai Si (eds.), CRC publishes: Boca Raton, Florida State (1974); The bioavailability of controlled drug (Controlled Drug Bioavailability), medicament production design and performance (DrugProduct Design and Performance), Smolen and Bauer (eds.), Wiley: New York (1984); Ranger and Peppas, polymer science periodical, the revised edition of polymer chemistry (J.Macromol.Sci.Rev.Macromol.Chem.) (1983) 23:61; Simultaneously referring to the profit dimension etc., science (Science) (1985) 228:190; During etc., neuroscience annual report (Ann.Neurol.) (1989) 25:351; Howard etc., Neurological Surgery periodical (J.Neurosurg.) (1989) 71:105).And in another embodiment, controlled release system can be placed on animal destination organization near, thereby only need body dose a part (as referring to, Goodson, in the medical applications of controlled release (Medical Applications ofControlled Release), on seeing, (1984) 2 volumes, 115-138 page or leaf).Especially, controllable release equipment can be incorporated in the animal body near unfavorable immune activation or tumour.(" science " be (1990) 249:1527-1533 (Science)) discusses by Lange in other controllable release systems in comment.

Provide following example to illustrate different situations of the present invention.In any case, the present invention can only not be confined to these examples.

Example 1:

Synthesizing of thiohydantoin derivative

General introduction: thiohydantoin derivative, or the compound of molecular formula (I), wherein R ₃Be group (a), differentiate by screening commercial available tryptophan derivative.N-methyl (R ₁=H; R _A=CH ₃R _B=H; X, Y, Z=H), N-allyl group (R ₁=H; R _A=-CH ₂CH=CH ₂R _B=H; X, Y, Z=H) and N-phenyl (R ₁=H; R _A=Ph; R _B=H; X, Y, Z=H).Proof thiohydantoin derivative is than distinct enzyme tryptophane 2 on the structure, and 3-dioxygenase (TDO2) all exceeds 50% on restraining effect and important selectivity.For further improving the potential of inhibitor, emphasis of the present invention may be exactly the replacement of indole ring, and this program has strengthened the potential of tryptophan derivative in the past.Particularly, reported literature points out that N-1, the C-5 of indole ring and the replacement of C-6 will provide more effective inhibitors (Cady and Sono (1991) biological chemistry and biophysics archives (Arch.Biochem.Biophys.), 291:326-33; Munn, D.H. etc. (1998) science (Science), 281:1191-3; Peterson, A.C. etc. (1994) medical chemistry research (Med.Chem.Res.), 3:531-544; Southan, M.D. etc. (1996) medical chemistry research (Med.Chem.Res.), 6:343-352).In addition, though do not report C-4 halide derivative (Southan etc.) in the literature, the present invention has determined 5-bromo-4-chloro-1, and the 3-diacetic acid esters is a kind of effective inhibitors (bigger by 50% than the restraining effect of IDO when 250 μ M).

Synthetic: as to synthesize thiohydantoin derivative (Fig. 1 by between indole-3-formaldehyde and thiohydantoin 1, carrying out Erlenmeyer-Plochl azlactone type condensation reaction; Djura and Fu Kena (1980) organic chemistry periodical (J.Org.Chem.), 45:735-7; Guella, G. etc. (1988) Helv.Chim.Acta, 71:773-82; Guella, G. etc. (1989) Helv.Chim.Acta, 72:1444-50; Daiweideke and Arthur D. Little (1959) chemical association periodical (J.Chem.Soc.), 729-731; Zhu Lian and Sturgis (1935) american chemical association periodical (J.Am.Chem.Soc.), 1126-28).Produce the mixture of conjugated alkene steric isomer 3 possibly, can detect these products is plane analogues of thiohydantoin.The reduction of conjugated alkene can be finished (for example, NaBH by the nucleophilic reductive agent in 3 ₄, Li (s-Bu) ₃BH and [(Ph ₃P) CuH] ₆), because the existence of sulphur can be forbidden more catalytic hydrogenation method commonly used.

Such as, can synthesize thiohydantoin (Fig. 1 by the reaction between for example glycine methyl ester or glycine ethyl ester and the alkyl isosulfocyanate; The west is covered and Ganesan (1997) organic chemistry periodical (J.Org.Chem.), 62:3230-35).Notably be, this thiohydantoin synthetic method allows various alkyl group to act on (Xi Meng and Ganesan (1997) organic chemistry periodical (J.Org.Chem.) on the N-1 nitrogen-atoms by reduction amination, 62:3230-35) and the N-3 nitrogen-atoms on combination, this depends on the selection of lsothiocyanates reagent.

On the basis of before IDO inhibitor research, with the condensation reaction of thiohydantoin in can use various halogen to replace indole-3-formaldehyde.5-bromo-and 5-chloro-indole-3-formaldehyde can obtain commercial.Can finish locational replacement (Fig. 2 of C-4 of indole-3-formaldehyde by the thallation-halogenation of indole-3-formaldehyde 5; Somei, M. etc., (1984) heterocyclic (Heterocycles), 22 (4): 797-801; Ohta, T. etc. (1987) heterocyclic (Hetereocycles) 26:2817-22; Somei, M. etc. (1985) chemical pharmacy bulletin (Chem.Pharm.Bull.) 33:3696-708).By with dimethyl carbonate react the N-that finishes indole-3-formaldehyde 6 methylate (Jiang, the deviation (Org.Proc.Res.Dev.) of the organic process study of X. etc. (2001), 5:604-8).

The generation of 6-halogenation base-indole-3-formaldehyde can by commercial be easy to utilize 2,4-dinitrotoluene (DNT) synthetic 6-amino indole synthesizes via the Leimgruber-Batcho indoles and carries out (Fig. 3; Batcho and Leimgruber (1990) organic synthesis set (Org.Syn.Coll.), 3:34).The 6-amino group can be used as ethanamide protection.Indole-3-formaldehyde 10 (Smith, G.F. (1954) chemical association periodical (J.Chem.Soc.), 3842-6 that the Wei Ersi Mel formylation reaction of C-3 position can provide 6-to replace; James and Snyder (1963) organic synthesis integrated (Org.Syn.Coll.), 4:539).The existence of the ethanamide group of electron release can improve the efficient of Wei Ersi Mel formylation reaction.Carry out being set forth before the methylating of indole nitrogen can resemble.Under acid reflux conditions, the 6-amino group can go protection, and amine 12 can transform into halogen (Somei, M. etc. (1985) chemical pharmacy bulletin (Chem.Pharm.Bull.), 33:3696-708 by Sandmeyer or uncommon door reaction then; Somei and Tsuchiya (1981) chemical pharmacy bulletin (Chem.Pharm.Bull.), 29:3145-57; Moriya, the Japanese chemical association's bulletins of T. etc. (1975) (Bull.Chem.Soc.Jpn.), 48:2217-8) or with peroxy acid oxidizing reaction generation nitryl group (Pagano and Emmons, (Org.Syn.Coll.) gathered in organic synthesis, 5:367).The side reaction of the 3-formaldehyde that relates in Sandmeyer, Xi Men or amine oxidizing reaction, this problem can solve by change the order of introducing 3-formaldehyde and methide on N-1 position after a while.

It is to finish (Fig. 4) by 14 electrophilic fragrance replacement that the synthetic of derivative of replacement all arranged on 5 and 6 positions.It is reported title, the nitrification of 1-methyl-indole-3-formaldehyde provide 5-nitro and 6-nitro mixture of products output 93% (Da Settimo and Saettone (1965) tetrahedron (Tetrahedron), 21:1923-9).The use of 6-acetamido derivative stops 6 locational nitration reactions, in addition, activates 5 positions and carries out electrophilic aromatic substitution.Except nitrification, bromination and chlorization also can be utilized, and institute is so that various 5, and the disubstituted indole-3-formaldehyde class of 6-is synthesized.In addition, 5, the thallation-halogenation of the disubstituted indole-3-formaldehyde 15 of 6-(Somei, M. etc. (1984) heterocyclic (Heterocycles), 22 (4): 797-801; Ohta, T. etc. (1987) heterocyclic (Heterocycles), 26:2817-22; Somei, M. etc. (1985) chemical pharmacy bulletin (Chem.Pharm.Bull.), 33:3696-708) the functionalization effect of permission C-4 position.And according to the method that elaborates in the method 3 (Fig. 3), C-4 ethanamide group can be modified.In conjunction with the time, these operations allow to obtain complicated polysubstituted indole-3-formaldehyde in a kind of mode efficiently.

It should be noted that, proof IDO has preference to the L-tryptophane, and (Peter is inferior, (1994) medical chemistry research (Med.Chem.Res.) such as A.C., 3:531-544), but also accept the D-tryptophane as a kind of substrate matter (Soho, M. (1989) biological chemistry (Biochemistry), 28:5400-7).Therefore, can not require a kind of body pure products that shines upon utterly.

Thiohydantoin derivative synthetic other method comprise 2 with condensation (Fig. 5 of the ethyl half ester of acetamidomalonic acid; Hengartner, U. etc. (1979) organic chemistry periodicals (J.Org.Chem.), 44:3741-7).Produce 19 with the hydrogenization of the catalyzer of Wilkinson (Horwell, D.C. etc. (1994) organic chemistry periodicals (J.Org.Chem.), 59:4418-23).The hydrolytic action of ethanamide and ethyl ester provides tryptophane analogue 20.Can provide thiohydantoin product (Cejpek, K. etc. (2000) agricultural food product chemical periodicals (J.Agric.Food.Chem.), 48:3560-5 with the reaction of alkyl lsothiocyanates; Mizrach and Polonska (1987) Khim.Farm.Zh., 21:322-9), may be more effective although on similar amino ester, carry out the lsothiocyanates reaction (Xi Meng and Ganesan (1997) organic chemistry periodical (J.Org.Chem.), 62:3230-35).

A potential important benefit of this selectivity approach is exactly to have the ability to obtain a kind of product that contracts that shines upon the body enrichment.Several asymmetric hydrogenization (Hengartner, U. etc. (1979) organic chemistry periodicals (J.Org.Chem.), the 44:3741-7 of the compound of relevant dehydrogenation tryptophane have been reported; Knowles, W.S. etc. (1975) american chemical association periodicals (J.Am.Chem.Soc.), 97:2567-8).So the metalepsy of the triphenyl phosphorus part of Wilkinson catalyzer and chirality phosphuret-(t)ed hydrogen part may cause the stereoselectivity of the L isomer of tryptophane analogue to be synthesized.

Example 2:

Synthesizing of Brassinin derivative

General introduction: prove that Brassinin and corresponding plants Hangzhoupro toxin have anti-cancer properties, it is reported it is that ((Cancer Metathesis Rev.), 21:231-255 are looked back in Parker and Pezzuto (2002) cancer metathesis for the prophylactic action of receiving from detoxifcation owing to carcinogenic substance; Mehta, R.G. etc. (1995) carcinogenesises (Carcinogenesis), 16:399-404; Mehta, R.G. etc. (1994) anticancer research (Anticancer Research), 14:1209-1213).Be enjoyably, brassilexin, a kind of relevant phytoalexin, it is reported that claiming is that (Peter is inferior for the noncompetitive inhibitor of IDO, (1993) medical chemistry research (Med.Chem.Res.) such as A.C., 3:473-482), yet, the work at initial stage demonstrates, and brassinin is the IDO competitive inhibitor.Till now, also there is not the brassinin derivative (that is: to have the compound of molecular formula (I), wherein a R ₃Be group (b)) or the brassilexin derivative (that is: have the compound of molecular formula (I), wherein a R ₃And R ₂In conjunction with forming (ii) or (iii)) the inhibiting correlative study of IDO.Brassinin and tryptophane structural similar and identify that it is a kind of competitive inhibitor thinks that derivatization is similar to the inhibitor on the tryptophane basis of having reported.

Synthetic: indole-3-formaldehyde discussed above (Fig. 2-4) can be used for producing the brassinin derivative, promptly has the IDO inhibitor of molecular formula (I), wherein R ₃Be group (b).Indole-3-formaldehyde 2 can change into 3-aminomethyl derivative 22 (Fig. 6) by carrying out reduction amination with azanol.3-aminomethyl indole derivatives potentially unstable (Kutschy, P. etc. (1991) Synlett, 289-290; Kutschy, (1998) Tet.54:3549-66 such as P.), so will handle with dithiocarbonic anhydride immediately, then handle with methyl iodide, generate brassinin derivative (Takasugi, Japanese chemical bulletin (Bull.Chem.Soc.Jpn.) 61:285 of association of M. etc. (1988) that replacement is arranged on the phenyl ring; Pedras, M.S.C. etc. (1992) life science progress (vegetable chemistry) (Life Science Advances (Phytochemistry)), 11:1; Mehta, R.G. etc. (1995) carcinogenesises (Carcinogenesis), 16 (2): 399-404).

Example 3:

Synthesizing of beta-carboline derivatives

General introduction: β-Ka Lin or carboline derivative promptly have the compound of molecular formula (I), wherein R ₃And R ₂In conjunction with forming (i), once reported (Sono and Cady (1989) biological chemistry (Biochemistry), 28:5392-5399 before; Peter is inferior, A.C. etc. (1993) medical chemistry research (Med.Chem.Res.), 3:473-482; Eguchi, (1984) biological chemistry such as N. and biophysics archives (Arch.Biochem.Biophys.), 232 (2): 602-609).But, do not report analogue---the 3-butyl-β-Ka Lin of the active derivative of tool.Particularly, β-Ka Lin nuclear the locational replacement of C-6 (F and-N=C=S) produce several active similar derivative things, do not test but combine the analogue that C-3 and C-6 replace; The also not test of replacement that the C-3 butyl of the β-Ka Lin that other any ratios have been studied is big (Peter is inferior, A.C. etc. (1993) medical chemistry research (Med.Chem.Res.), 3:473-482).At C-5, the beta-carboline derivatives that replaces on 7 and 8 positions is not reported yet.Allowing the people interested especially is that the beta-carboline derivatives of report is as the mechanism of action of competitive inhibitor, for example, on the heme iron of iron enzyme with oxygen competition-individual binding site (Sono and Cady (1989) biological chemistry (Biochemistry), 28:5392-5399).

Synthetic: the coml practicality of β-Ka Lin-3-carboxylic acid ethyl ester 24 makes the chemical variation of this structure have synthetic (Fig. 7) that enough magnetisms are used for beta-carboline derivatives.The nitrification of 24 C-6 position (Settimj, G. etc. (1988) heterocyclic chemistry periodical (J.Heterocyclic Chem.), 25:1391-7) and iodization (Huth, A. etc. (1987) Tet. 43:1071-4) has realized good yield.In addition, bromination (Pang Sai and Erra-Balsells (2001) heterocyclic chemistry periodical (J.Heterocyclic Chem.), the 38:1087-1095 of the selection of the regionality of β-Ka Lin; Kardono, L.B.S. etc. (1991) natural product periodicals (J.Nat.Prod.) 54:1360-7) and chlorization (40:419-426) process is also reported for Ponce, M.A. etc. (2003) heterocyclic chemistry periodical (J.Heterocycllic Chem.).The reduction of C-6 nitration product realizes (Settimj, (1988) heterocyclic chemistry periodical (J.Heterocyclic Chem.) such as G., 25:1391-7), the amine that the result produced has been used for the locational direct bromination (Huth of C-5 of β-Ka Lin, (1987) Tet. such as A., 43:1071-4).These reactions will promote the synthetic of beta-carboline derivatives that C-5 and C-6 replace.

The locational derivatization of C-3 can realize (Fig. 8) via C-3 formaldehyde.24 C-3 ester and the chemical selective reduction of the DIBAL oxygenizement of ethanol and chemo-selective (or be reduced into) can produce C-3 formaldehyde.Organolithium or Grignard reagent add in the acetaldehyde can produce alcohol derivate 29.The dehydration of this alcohol can produce alkene 30, and it can optionally reduce and produce the C-3 alkyl group of different lengths, and this depends on selecting for use of organometallic reagent.Also can test pure and mild alkene intermediate.The two can select one, can produce alkene with the Wittig reaction of formaldehyde.

The other method of beta-carboline derivatives relates to cyclic action and oxygenizement (Fig. 9 of tryptophan derivative 32; Haffer, G. etc. (1998) heterocyclic (Heterocycles) 48 (5): 993-8).The previous tryptophan derivative of being discussed (Fig. 2-4) can carry out cyclization and form pyrido-indole structure.An advantage using this operation is exactly the I D0 inhibitor of having the ability to produce on the potential β-Ka Lin basis, and carries out the functionalization effect on the C-7 position of β-Ka Lin.

Example 4:

Synthesizing of 3-amino-2-naphthoic acid derivative

General introduction: 1994, inferior and the colleague of Peter has reported that 3-amino-2-naphthoic acid is effective I DO inhibitor (Peter is inferior, A.C. etc. (1994) medical chemistry research (Med.Chem.Res.) 3:531-544) among tryptophan derivative and related compound.It should be noted that, in article, do not report other derivatives of 3-amino-2-naphthoic acid, according to known at present, there is not the report of the structural trial of this related derivatives, although in fact this compound is more effective than prototype IDO inhibitor L-1-methyl-tryptophane (1MT) yet.Since 3-amino-naphthoic acid is-kind of competitive inhibitor that (being similar to C-4, C-5 and the C-6 position of the indole ring of tryptophane) all has substituent derivative that a kind of better IDO inhibitor can be provided on C-6, the C-7 of naphthalene nuclear and the C-8 position.

Synthetic: as, on two rings, all to have naphthalene class synthetic of substituent asymmetric replacement to be still a challenge though exist a lot of methods to synthesize naphthalene compounds (de Koning, C.B. etc. (2003) Tet.59:7-36).Yet the phenyl ring reaction of the terephthal aldehyde that the first step replaces can produce 3-amino-2-naphthoic acid (Kienzle, F. (1980) the Helv.Chim.Acta 63:2364-9 of replacement; Singh and Khade (2002) bio-link chemistry (Bioconj.Chem.) 13:1286-91).The synthetic fast validity (Figure 10) that has proved this method of 3-amino-7-bromo-2-naphthoic acid 36.

Can synthesize terephthal aldehyde 34 by 4-bromo-oxygen-dimethylbenzene or dimethyl phthalic ester.Within a step, just can synthesize ethyl-3-nitropropionic acid ester (Belikov, V.M. (1956) Izv.Akad.Nauk., S.S.S.R. by silver nitrite and ethyl-3-ester bromopropionylamino; Otdel.Khim.Nauk 855-62; Chemistry summary (Chemicalabstract) (1957) 51:1837j).The condensation reaction meeting of these two kinds of compounds produces the mixture 35 of 6-bromo and 7-bromo 3-nitro-2-naphthoic acid.The reduction reaction of nitryl group and the hydrolytic action of ester group can produce the 3-amino-2-naphthoic acid of bromination.These two kinds of regional isomers can separate at any point in synthetic, but these two kinds of compounds can test in a kind of mode fast, two kinds of compounds are fitted in together probably in this mode, surpass any inconvenience of the mixture of regional isomery.

The synthetic regionality of considering of the 3-amino of other replacements-2-naphthoic acid class is controlled, and replaces bromine (Figure 11) by using electron-donating group.O-phthalaldehyde(OPA) 37 can be by 3, and 4-xylidine or dimethyl phthalic ester generate.Amino electrophilic aromatic series replaces or acetamido terephthal aldehyde precursor allows group to be incorporated on the C-6 position of 3-amino-2-naphthoic acid.Because the existence of the acetamido group of electron release, so the phenyl ring reaction is regional select (Kienzle, F. (1980) Helv.Chim.Acta 63:2364-9).After the phenyl ring reaction, 7-acetamido or amino group are considered via the substitution reaction of electrophilic fragrance in the locational effective replacement of C-8.Go after the protection, the 7-amino group can or be wished the door reaction through diazotization and Sandneye and be produced various substituting groups.Nitryl group reduction and hydrolysis of ester group can produce the derivative of 3-amino-2-naphthoic acid, the compound that contains molecular formula (II) that promptly is used to test.

As another kind of method, methoxyl group o-phthalaldehyde(OPA) 43 (Z=H) with the condensation reaction of 3-nitropropionic acid ester in shown sizable control (Figure 12; Kienzle, F. (1980) Helv.Chim.Acta 63:2364-9).Methoxyl group instructs and carry out the aromatic substitution reaction of electrophilic on the position, ortho position, just methoxyl group is changed into amine changes into halogen then, can utilize method (Wu Erfu, J.P. etc. (2000) organic chemistry periodicals (J.Org.Chem.) 65:1158-74 of Buchwald-Hartwig ammonification; Wu Erfu and Buchwald (1997) organic chemistry periodical (J.Org.Chem.) 62:1264-7; The Louis, J. etc. (1997) organic chemistry periodicals (J.Org.Chem.) 62:1268-73; Hartwig, J.F. (1998) Angew.Chem.Int.Ed.Engl.37:2046-67).Methoxyl group changes into trifluoro thing 45 and considers ammonification, but carboxylicesters is because strong alkaline condition and process is incompatible therewith.Therefore, 3-nitro-propionitrile---can be similar to ethyl 3-nitro propionic ester and synthesize just and will can use in condensation reaction with 3-bromo propionitrile as parent material, cyano group can be hydrolyzed in end of synthesis.

Example 5:

Synthesizing of Brassilexin derivative

The Brassilexin derivative, the compound of molecular formula (I), wherein R ₃And R ₂In conjunction with form (ii) or (iii), can synthesize by method shown in Figure 13 (as, referring to Devys and Barbier (1993) Org.Prep.Proc.Int.25:344-346).

The synthetic schemes of the brasilexin derivative that replaces shown in Figure 14 and 15 (as, referring to (2002) Tet.Lett.43:5793-5795 such as Yeung).

Example 6:

Synthesizing of cyclopropyl/'-aziridino tryptophan derivative

The scheme of synthetic cyclopropyl/'-aziridino tryptophan derivative, R ₃Be the compound of (c) or molecular formula (d) (I), shown in Figure 16 and 17 (as, referring to (1996) tetrahedron (Tetrahedron) 52:9901-9908 such as Donati; Ishikawa etc. (2001) american chemical association periodicals (J.Am.Chem.Soc.) 123:7705-7706).

Example 7:

Synthesizing of the competitiveness of mooring/noncompetitive derivative

Figure 18 demonstrates competitiveness/noncompetitive derivative, the i.e. R of synthetic mooring ₃It is the synthetic schemes of compound of the molecular formula (I) of (e).

Example 8:

Synthesizing of two dehydrogenation derivatives

Fig. 1 shows the synthetic method of two dehydrogenation derivatives (compound 3 among Fig. 1), i.e. R ₃It is the compound of the molecular formula (I) of (f).Example 1 above provides relevant these compound more detailed descriptions.Particularly, the diastereomer that also comprises E and Z among the present invention.

Example 9:

The mensuration of Novel IDO inhibitor

1. the biochemical measurement of Novel IDO inhibitor:

General introduction: the biological chemistry of IDO is fine to be determined, is to separate a kind of enzyme (Higuchi, K. etc. (1963) federal journal (Federation Proc.) 22:243 (summary) ((abstr.)) of coming out at first in 1963; Shimizu, T. etc. (1978) biological chemistry periodicals (J.Biol.Chem.) 253:4700-6).IDO is a kind of molecular weight single-unit of being approximately 41kDa, the oxydo-reductase that contains protoheme.For keep the active ferrous iron form in external katalysis, enzyme require methylene blue and superoxide or reductive agent are as the xitix combination.In vivo, think that four hydroxyl keto-alcohols or tetrahydrobiopterin can realize the effect of methylene blue dye, be in probably on the special position of noncompetitive IDO inhibitor.Organized enzyme can by mammalian genes in bacterium, demonstrate the clone, His mark form produces (Littlejohn, T.K. etc. (2000) Prot.Exp.Purif.19:22-29).This also provides a kind of data source easily for the biochemical analysis of enzyme.The active conventional biochemical assay of IDO (Daubener, W. etc. (1994) immunological method periodicals (J.Immunol.Methods) 168:39-47) of carrying out on the spectrophotometry basis of the kynurenine (hydrolysate of N-formyloxy-kynurenine) that produces from tryptophane is used as the readout of chemically examining on enzyme and the cell base.The chemical examination of enzyme for discriminating have IDO suppress active compound provide a kind of easily, the examination of high throughput.This chemical examination also is used to measure the Ki value of special compound, and this is worth the development of the structure-activity relationship (SAR) that centers on different series of compounds very important.Chemical examination on the cell base can determine that the IDO of the compound differentiated suppresses active, also can propose bioavailability---compound suppresses the initial results of the ability of IDO in the cell.The inhibiting singularity of IDO by with other known tryptophanes contrast chemical examinations on cell base that enzyme tryptophane dioxygenase (TDO is also referred to as TDO2 in the literature) is decomposed in check.

Method: the cDNA clone of the IDO of people and mouse separates sequentially and checks.For preparation is used for the enzyme of biochemical research, the IDO protein of the His mark of C-end can utilize IPTG inductive pET5a vehicle system to produce, and separate on the nickel post in large intestine Ai Shelixishi bacillus.Can contrast the proteinic turnout that the concentration of estimating is come the verification portion purifying by gel electrophoresis and with the protein standard.In order to chemically examine the enzymic activity of IDO, can be according to published operation (Littlejohn, T.K. etc. (2000) Prot.Exp.Purif.19:22-29; Takikawa, O. etc. (1988) biological chemistry periodicals (J.Biol.Chem.) 263:2041-8) carrying out 96 used orifice plate spectrophotometrys of kynurenine product experimentizes.For differentiating that IDO suppresses active evidence,, measure the compound of single concentration in 100 μ l reaction volumes as 200 μ M with the tryptophane contrast that the IDO enzyme of 50ng increases progressively with the concentration of adding as 0,2,20 and 200 μ M.The kynurenine product can be measured in the time of 1 hour.Can collect the compound that enzyme dynamic date widely is used to select.Determining as shown in figure 19 of the optimum Ki value of 1MT (Ki=34.6 μ M) and MTH-Trp (Ki=11.4 μ M).These data presentation go out, and the thiohydantoin form directly suppresses IDO enzymic activity and potential than about 3 times of the potential that obtains with 1MT.

Following process is the example of the experiment on the cell base.Recommend to utilize fat transfection amine 2000 reagent (Invitrogen) by manufacturers, the COS-1 cell carries out transient transfection with the plastid of the CMV promotor driving of the cDNA that shows IDO.One association's cell carries out transient transfection by the plastid that TDO shows.After the transfection 48 hours, cell was allocated to 96 hole forms, and every hole has 6 * 10 ⁴Individual cell.Second day, wash these holes, add the new medium (phenol red radical) that contains 20 μ g/ml tryptophanes with inhibitor.Reaction is carried out stopping after 5 hours, removes supernatant liquor, by the method for the enzyme quantitative analysis of being set forth kynurenine is carried out spectrophotometry experiment (Littlejohn, T.K. etc. (2000) Prot.Exp.Purif.19:22-29; Takikawa, O. etc. (1988) biological chemistry periodicals (J.Biol.Chem.) 263:2041-8).Be to obtain the active elementary affirmation of IDO, compound can be measured during as 100 μ M in single concentration.The compound that the concise and to the point description that can collect dosage widely increases is used to select.The determining of EC50 value that shows 1MT (EC50=267 μ M) and MTH-Trp (EC50=12.9 μ M) among Figure 20.These data show that as for suppressing intracellular IDO, MTH-Trp is than 1MT more much effective (about 20 times).

2. drug effect/the medicine of Novel IDO inhibitor is moving measures:

General introduction: the intraperitoneal of bacteria lipopolysaccharide (LPS) understand bring out the IDO activity in various tissue, causes the generation of kynurenine, and is discharged into blood flow interior (Figure 21).LPS uses and just reaches the highest kynurenine level (Takikawa, O. etc. (1986) biological chemistry periodicals (J.Biol.Chem.) 261:3648-53 one day after; Yoshida, H. etc. (1998) cells (Cell) 94:739-750).Drug effect analysis as described herein is on the basis of the serum level of measuring kynurenine and tryptophane.Calculate kynurenine/tryptophane ratio and can estimate the activity of IDO, this activity does not rely on basic tryptophan levels (Fuchs, D. etc. (1991) immunology communications (Immunol.Lett.) 28:207-11; Gasse, European periodical (Eur.J.Clin.Chem.Clin.Biochem.) 32:685-9 of (1994) clinical chemistry such as T. and clinical biochemistry), the active method of this measurement IDO is used for the mankind continually.This method is that it is a kind of Noninvasive process and allows to gather several samples from same animal to the principle advantage of IDO enzymic activity in the direct assessment tissue.This can monitor the activity of IDO on a plurality of time points in a mouse body.Tryptophane in the serum and kynurenine level can be analyzed to determine by HPLC.Compound level in the serum also can be measured by identical high performance liquid chromatograph, allows to collect simultaneously in a test the moving data of medicine like this.

Method: can utilize about 8-10 week big FVB MMTVneu male mouse to carry out most drug effect analysis because their genetic background be used in the mammary cancer test in to assess the female mouse of MMTVneu of compound efficacy identical.The salmonella Minnesota State mutant R-595 (S.minuetsota R) that stems from LPS demonstrates, in the comparative analysis of the LPS preparation of different bacterium kind strain, can elicit the kynurenine fall out effect (Yoshida, (1981) biological chemistry such as R. and biophysics archives (Arch.Biochem.Biophys.) 212:629-37) of the level that delays most.Because the influence power that it can provide the wideest window to evaluate the IDO inhibitor, this LPS preparation can be used in the drug effect analysis.LPS Sha Shi Minnesota R can elicit the active minimum i.p. bolus injection dosage of the highest IDO, reported it is about 1mg/kg, LPS treatment back is in the time of about 24 hours, the active function of IDO reaches the highest (Yoshida, (1981) biological chemistry such as R. and biophysics archives (Arch.Biochem.Biophys.) 212:629-37).

The serum level of kynurenine and tryptophane can be analyzed by HPLC and quantitatively determine (Hwu, P. etc. (2000) immunology periodical (J Immunol) 164:3596-9; Widner, B. etc. (1997) clinical chemistries (Clin.Chem.) 43:2424-6; Referring to Figure 22 A-D).By this process, detect the serum-concentration of at least 1.25 μ M kynurenines and 3 μ M tryptophanes.In the male mouse body of the FVB that does not stimulate, the serum kynurenine is on the detection limit value or is lower than limit value, and the serum tryptophane is easy to just can detect (Figure 22 B) when 50 μ M.LPS excites back 24 hours, and the serum kynurenine brings out to about 6 μ M (Figure 22 C).1MT also can measure when at least 5 μ M concentration effectively in the serum.This just is under the serum level of 1MT of the about 100 μ M that biologically effectively reach during dosage of 1MT piller of 2 * 140mg (Figure 22 D).

At first excite the compound that when the serum kynurenine reaches maintenance level, gives bolus injection dosage then, can assess this compound with LPS.The kynurenine pond is along with (Bender and McCreanor (1982) Acta Biochimica et Biophysica Sinica (Biochim.Biophys.Acta) 717:56-60 that overturns rapidly less than the transformation period in 10 minutes the serum; Takikawa, O. etc. (1986) biological chemistry periodical 261:3648-53), exceedingly do not cover IDO restraining effect to the kynurenine product so do not expect the kynurenine that pre-exists.The carrier that selection is used for administration can depend on the physical properties of each particular compound to a great extent.Optimum carrier is an isotonic saline solution, but this needs compound to dissolve in the aqueous solution.Can reckon with that some compounds may not can fully dissolve, in this case, compound can be used as methylcellulose gum ^/ tween ^Suspension in (0.5% methylcellulose gum/1% tween  80) is used.

Every experiment all may comprise the mouse (for the IDO activation provides clear and definite control) that the mouse (determine original kynurenine level, compare with the another mouse) of non-LPS contact and the thing that only utilizes the medium that one group of LPS contacts are taken medicine.Mouse can be monitored after taking LPS, just implements euthanasia (hair is holded up, and is dull) immediately if they present significant endotoxemia sign.Every kind of compound can be measured in the mouse body of single high i.p. bolus injection dosed administration in the 100mg/kg scope at least at first.Can (for example gather blood by the timed interval of definition, taking behind the compound and to get 50 μ l/ samples in 5,15,30 minutes and 1,2,4,6,8 and 24 hour) HPLC that is used for kynurenine and tryptophan levels analyzes (drug effect analysis), also is used for the analysis (medicine is moving to be analyzed) of compound level simultaneously.From the moving data of medicine, can measure the maximum serum concentration that compound reaches and the estimating speed of removing.The level of compound in serum by different time points relatively be with respect to kynurenine/tryptophane ratio, roughly the inhibiting effective IC of IDO in the estimated body ₅₀

On the basis of single dose research, each compounds effective can carry out the dosage increase research second time.For example, research reaches the inhibiting maximal dose of 100%IDO in the time of can be at the inherent peak concentration of one group of mouse (if possible), gradually reduces to cover 2 log between maximum dose level and the lowest dose level for other several groups concentration of taking medicine with 3 multiplication amounts ₁₀Scope.Can from these data, choose accurate IC ₅₀Measured value.Same method can be used for the oral bioavailability of test organisms active compound, at first test each compound when single peak concentration p.o. bolus dosage, further being determined at dosage then increases those compounds that show significant oral effect in the research.For guarantee to respond in the body influence of the dimorphism that is not subjected to sex, the IC that single i.p bolus injection dosage experiments can be calculated with every kind of active compound in female mouse ₅₀Carry out on the dosage.

Example 10:

Utilize the tumour combined therapy of IDO inhibitor and signal transduction inhibitor

" tumor mouse " test pattern is measured IDO inhibitor and the effect of STIs on tumor pathophysiology to utilize the MMTVneu of breast cancer to transcribe.MMTVneu transcribes mouse makes mammary gland aggressiveness tumour become big, is similar to the human body breast ductal cancer that is not easily distinguishable.In MMTVneu mouse test pattern, breast cancer is caused by the tissue specific expression of the HER-2/neu transgenation kind that frequently is activated in aggressiveness human body breast ductal cancer.HER-2 is an one's share of expenses for a joint undertaking of the EGF-R section of cell surface growth factor receptors.Myc is the downstream effector that a kind of HER-2/Neu drives the obligate of cancer.Female MMTVneu " tumor mouse " mating impels for twice mammary tumor virus (MMTV) promotor (Neu/HER2 oncogene transcribes in the driving mammary tissue) of mouse to express.In this test macro＞90% mouse produced breast tumor when big or small in 5 months.Has about 150mm ³The MMTVneu of similar size tumor " tumor mouse " is random assignment contrast or experimental therapy group.The contrast mouse is implanted placebo and discharges piller (innovation research company limited, Sarasota, Florida State) in limited time.Mouse experiment group (1) implant contain 1MT discharge piller in limited time, (2) use L-744,832 treatments, or (3) implant and contain discharging piller in limited time and using L-744 of 1MT, 832 treat.L-744,832, exactly like the CaaX motif that adds farnesyl, be a kind of effectively, farnesyl tranfering enzyme (FTI) inhibitor (Kohl etc. (1995) country medicine (Nat Med.) 1 (8): 747-748) optionally.

Discharge piller in limited time and be made of a kind of multipolymer, this multipolymer is an inert, can progressively dissolve and resolve into non-toxic substance, and this material mainly is retained in a certain part in experimentation.Can 10mg/ days dosage (innovation research company limited/Innovative Research, Inc., Sarasota, Florida State) of release in 14 days with the piller that discharges in limited time that 1MT injects by the supporting documents of commercial supplier.It is 20mg/ days that every mouse is implanted two pillers release total doses.Therefore, for the mouse of a 25g, total dose is 800mg/kg/ days or 280mg in 14 days.Based on the specification sheets of manufacturer, reach steady-state level in 12-24 hour, and in whole period, keep this level.Press people's such as Munn elaboration (science (Science) 281:1191-1193,1998), the dosage of release is effective (A. bridle, J.B.DuHadaway, G.C.Prendergast, the result who does not deliver) to the rejection that elicits allochthonous conceptus.

Discharge piller in limited time in the subcutaneous implantation of mouse back of anaesthetizing with ketamine/xylazine intramuscular injection.Use follows the blunt dissection of mosquito forceps that skin is separated to produce subcutaneous satchel from underlying muscle.One or two biodegradable slowly-releasing piller is implanted in this satchel, rather than directly below otch, so that prevent mechanical stress and wound dehiscence.With wound clamp otch is closed up then.Discharge piller in limited time and support conceivedly on the capability foundation discussion of normal intrapartum female mouse implanting placebo, as if can ignore by the misery that this process produces.

Signal transduction inhibitor, FTI L-744 for example, 832, press people's such as Kohl elaboration, prepare and offer mouse (natural medical science (Nature Med.) (1995) 1:792-797).

Figure 23 has summarized test 1MT and FTI L-744, and 832 (also being chemotherapeutics) cause the result of study of the ability experiment of set tumour regression in MMTVneu " tumor mouse " pattern together.In the experimentation of fortnight, the gross tumor volume of observing simulation treatment contrast mouse has increased about 200%.With 20mg/ days 1MT treatment mouse, provide by the subcutaneous piller that discharges in limited time, tumour is slowed down but do not stop its growth.Similarly, suffer from the mouse of tumour and use FTI L-744,832 treat and can slow down but can not stop growth of tumor.On the contrary, 1MT adds L-744 in this test pattern, and 832 combined therapy can make tumour regression.

Example 11:

The test Novel IDO inhibitor

Screen various compounds and determine its effect as the IDO inhibitor.Some compound screens by following biochemical assay.IDO cDNAs is expressed as the His labelled protein in bacterium, carry out purifying (Littlejohn etc. (2000) experiment purifying magazine (Prot.Exp.Purif.) 19:22-29) by described mode before.In brief, pure IDO hatches and changes the quantity of IDO inhibitor trial-product with substratum.The fluorescence of measuring reaction mixture to be determining the effect for the examination inhibitor because reaction product---kynurenine is a fluorescence.External biological chemistry results of screening is set forth in Figure 24.

Test compound also screens (as for similarly testing (1999) J.Exp.Med.189:1363-1372 such as please refer to Munn) in the chemical examination on cell base.In brief, human body 293/ Phoenix cell is expressed the vector transient transfection by human body IDO or TDO cDNA.Test compound adds on the cells transfected with different concentration.Utilize method on the fluorescence basis in tissue culture medium (TCM), to estimate the quantity of kynurenine.Show these result of experiment among Figure 25-27.

By the indicated result of these figure, the effective inhibitors of having differentiated is the thiohydantoin derivative of one group of indoles amine.Figure 26 provides the result who uses these specific inhibitors.The most effective in these inhibitor---methyl-TH-DL-trp, the active restraining effect of IDO that demonstrates when concentration is 250 μ M is than 1MT high 2.7 times (Figure 27).

Except the thiohydantoin derivative of indoles amine, also filter out one group of natural product.Be that effective inhibitor of this group product comes from the compound (for example brassicaceous vegetable) of the food with anti-cancer characteristic enjoyably.Brassinin---a kind of compound of in Chinese cabbage, finding have with the natural product that is defined as the IDO inhibitor in the same effect (Figure 25 A) of compounds effective.

The toxicity of the compound that some screen also will detect.As shown in figure 28, most of IDO inhibition compounds are not just to have growth-inhibiting or cytotoxic (Figure 28) in itself to mammary gland or the prostatic cell that surgical plastic transforms.

Example 12:

The tumour combination treatment of IDO inhibitor and cytotoxicity chemotherapeutics

MMTVneu transgenosis " tumor mouse " model of mammary cancer also is used to measure the influence to tumor pathophysiology of IDO restraining effect and cytotoxin chemotherapeutics.

Has about 150mm ³The MMTVneu of similar big or small tumour " tumor mouse " is random assignment contrast or experimental therapy group.The contrast mouse is implanted placebo and discharges piller (innovation research company limited/Innovative Research, Inc., Sarasota, Florida State) in limited time.Mouse experiment group (1) implant contain 1MT discharge piller in limited time, described in example 10, (2) with taxol (taxol ) or the treatment of other cytotoxic agents, or (3) implantation contains discharging piller in limited time and using taxol (taxol ) or the treatment of other cytotoxic agents of 1MT.

Discharge piller by described before in limited time in the subcutaneous implantation of mouse back.

Prepare the cytotoxin chemotherapeutic as follows and discharge and give mouse.The solubilizing agent Cremophor that taxol is dissolved in isopyknic raw spirit and uses clinically ^Among the EL.Solution is preserved a week as the standing solution of 20mg/ml through sonic treatment 30 minutes in the time of 4 ℃.Before the use, this solution will further dilute with stroke-physiological saline solution with 1: 5 ratio.She Ji taxol is expelled in the tail vein by single bolus injection vein (i.v.) to the mouse clothes down in this way.Mouse tail can heat to be convenient to identify and intravenous injection.The maximum tolerated dose of taxol (MTD) (13.3mg/kg) is injected five times at 2 all experimental sessions of three time-of-week tables, and (that is Friday,---piller is implanted; Monday/Wednesday/Friday, Monday/Wednesday---taxol injection; Friday---allow animal euthanasia and obtain tumour and be used for analyzing).According to clinical preparation in the salt solution and by the bolus injection intravenous injection of same timetable, obtain MTD (1mg/kg) along chlorine ammonia cypress.Contrast treatment mouse receives the Cremophor that does not have taxol ^EL media formulating of recipe.

Figure 29 and table 1 add Figure 23, have summed up in MMTVneu " tumor mouse " pattern test 1MT and have caused the experimental result of the degeneration ability of set tumour with two kinds of cytotoxic agents.During the experimentation in two weeks, the gross tumor volume of observing simulation treatment contrast mouse has increased about 200%.With 20mg/ days 1MT treatment mouse, provide by the subcutaneous piller that discharges in limited time, tumour is slowed down but do not stop its growth.Similarly, the mouse that suffers from tumour is treated by intravenous injection with the taxol of maximum tolerated dose or suitable chlorine ammonia cypress and can slow down but can not stop tumor growth.On the contrary, 1MT adds that the combined therapy of taxol or suitable chlorine ammonia cypress can make tumour regression in this test model.Taxol decrement to 25% o'clock of about maximum tolerated dose can be observed similar result (not display data).Because the cytotoxic agent that uses in these researchs is deleterious to the T cell that really will recover and activate the IDO inhibitor as everyone knows, thus in view of these results of application before all be unexpected.

	Untreated	1MT only	Taxol only	The 1MT+ taxol	Only along chlorine ammonia cypress	1MT+ is along chlorine ammonia cypress
							Mouse quantity
	5	5	5	6	3	3
							Tumour quantity	5	7	6	9	5	5
Mean value	195.1	80.27	139.4	-30.2	91.35	-27.94
							Standard deviation	97.54	73.12	118.1	30.7	118.5	35.1
Standard error	43.62	27.64	48.2	10.23	53	15.7
							Minimum value	122.2	0	20	-78.4	-26.53	-67.86
25% percentage point		25	40.25	-56.5
							Intermediate value	134.4	72.87	130.4	-23.44	40	-30.56
75% percentage point		130.4	247.6	-6.445
							Maximum value	336.5	215	360.8	12.5	255.6	14.29
The 95%CI lower limit	73.95	12.65	15.52	-53.8	-55.79	-71.52
							The 95%CI upper limit	316.2	147.9	263.3	-6.605	238.5	15.64

Table 1

After the different therapies, the statistical study of the tumour of MMTVneu mouse.The gross tumor volume correlated percent change preceding with gross tumor volume after the treatment and treatment provides numeral.The lower limit of 95%Cl and the upper limit are represented the lower value and the higher limit of 95% cofidence limit.

The histology of isolating tumor biopsy and immunohistochemical analysis have shown the theatrical variation on the tumor tissues that only occurs in the mouse for the treatment of with combined method from control group and experimental group.Be the most significantly, in mouse body, found with combined method treatment the CD3 positive T cell significantly hemorrhage, cell dead and the relevant testimony (not display data) of infiltration.In a word, to degenerate be effective to the applied in any combination of 1MT and the cytotoxic agent breast tumor of bringing out regulation in MMTVneu " tumor mouse " model system.

Though above described and for example clear clearly several preferred implementations of the present invention, and do not meant that the present invention can be confined to these situations.May also do various corrections in addition and not violate scope and spirit of the present invention, as what proposed in the following statement.

In order to set forth the situation of the technical field of the invention more fully, several publications and patent document in explanation before, have been quoted.These citing documents content separately are hereby incorporated by.

Claims (37)

1. have indoleamine 2,3-dioxygenase (IDO) suppresses active compound, and this compound has the molecular formula of selecting from formula (I) and formula (II):

Divide formula (I):

R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups: R wherein _AAnd R _BFrom group H and alkyl, select independently; R wherein _CFrom group H and alkyl, select; Wherein n is from 1 to 10 integer, R _DIt is molecular formula

The carboline substituting group; With R wherein _AAnd R _BBe respectively from group H and alkyl, to select; Or R ₂And R ₃Combine and representative formula (I) partly concoct with the pyrroles and from group: With

The part of the middle ring of selecting, R in the base _EBe alkyl or alkyl-Q, Q represents molecular formula

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; With work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II):

Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

2. contain claim 1 compound of significant quantity and the pharmaceutical composition that is used for the treatment of cancer that pharmacy can be accepted the allowable carrier media.

3. one kind is the patient treatment method for cancer that needs it, comprise and use at least a indoleamine 2 of containing of significant quantity, and the pharmaceutical composition of 3-dioxygenase (IDO) inhibitor, at least a IDO inhibitor is selected from the above-mentioned compound that molecular formula (I) arranged:

Molecular formula (I) R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups: R wherein _AAnd R _BFrom group H and alkyl, select independently; R wherein _CFrom group H and alkyl, select; Wherein n is from 1 to 10 integer, and R _DIt is molecular formula

The carboline substituting group; With R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group:

The part of the middle ring of selecting, R in the base _EBe alkyl or alkyl-Q, Q represents molecular formula Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

4. in the method for claim 3, above-mentioned cancer is to select from the cancer of following histoorgan: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), an endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

5. in its patient of needs, treat method for cancer for one kind, comprising side by side or according to priority uses at least a indoleamine 2 of certain significant quantity for above-mentioned patient, 3-dioxygenase (IDO) inhibitor and at least a signal transduction inhibitor (STI), wherein, above-mentioned at least a IDO inhibitor is to select from one group of compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups:

R wherein _AAnd R _BFrom group H and alkyl, select independently;

R wherein _CFrom group H and alkyl, select;

Wherein n is from 1 to 10 integer, R _DIt is molecular formula

The carboline substituting group; With R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group:

With The part of the middle ring of selecting, wherein R _EBe alkyl or alkyl-Q, Q represents molecular formula

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

6. in the method for claim 5, above-mentioned at least a STI is from being to select the inhibitor in kinases or Akt path and the cell cycle kinase inhibitors by bcr/abl kinase inhibitor, Urogastron (EGF) acceptor inhibitor, her-2/neu acceptor inhibitor, farnesyl tranfering enzyme inhibitor (FTIs), Akt.

7. in the method for claim 6, wherein above-mentioned at least a STI is from by STI571, SSI-774, C225, ABX-EGF, Herceptin, L-744,832, select among rapamycin, LY294002, flavopiridal and the UNC-01.

8. in the method for claim 7, wherein above-mentioned at least a STI is L-744,832.

9. in the method for claim 5, wherein above-mentioned at least a IDO inhibitor and at least a STI can use simultaneously.

10. in the method for claim 5, wherein above-mentioned at least a IDO inhibitor and at least a STI can use in order.

11. in the method for claim 10, wherein above-mentioned at least a IDO inhibitor was used before at least a STI.

12. in the method for claim 10, wherein above-mentioned at least a STI used before at least a IDO inhibitor.

13. in the method for claim 5, wherein above-mentioned cancer is to select from the cancer of following histoorgan: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), an endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

14. pharmaceutical composition that is used for the treatment of cancer, said composition contains at least a indoleamine 2 of certain significant quantity in the pharmaceutical acceptable carrier medium, 3-dioxygenase (IDO) inhibitor and at least a signal transduction inhibitor (STI), wherein above-mentioned at least a IDO inhibitor are to select from the compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H;

R ₃By selecting in the following groups: R wherein _AAnd R _BIndependently

From group H and alkyl, select; R wherein _CFrom group H and alkyl, select;

Wherein n is from 1 to 10 integer, R _DIt is molecular formula The carboline substituting group; With

R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group With The part of the middle ring of selecting, wherein R in the base _EBe alkyl or alkyl-Q, Q represents molecular formula

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

In the molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

15. in the pharmaceutical cpd of claim 14, wherein above-mentioned at least a STI is from being to select the inhibitor in kinases or Akt path and the cell cycle kinase inhibitors by bcr/abl kinase inhibitor, Urogastron (EGF) acceptor inhibitor, her-2/neu acceptor inhibitor, farnesyl tranfering enzyme inhibitor (FTIs), Akt.

16. in the pharmaceutical cpd of claim 15, wherein above-mentioned at least a STI is from by STI 571, SSI-774, C225, ABX-EGF, Herceptin, L-744,832, select among rapamycin, LY294002, flavopiridal and the UNC-01.

17. in the pharmaceutical cpd of claim 16, wherein above-mentioned at least a STI is L-744,832.

18. the method for a treatment chronic viral infection in its patient of needs, comprise and use certain effectively at least a indoleamine 2 of quantity side by side or according to priority for above-mentioned patient, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutics, wherein above-mentioned at least a IDO inhibitor are to select from one group of compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups:

R wherein _AAnd R _BFrom group H and alkyl, select independently; R wherein _CFrom group H and alkyl, select; Wherein n is from 1 to 10 integer, R _DIt is molecular formula The carboline substituting group; With R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group

With

The part of the middle ring of selecting, wherein R in the base _EBe alkyl or alkyl-Q, Q represents molecular formula Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

19. in the method for claim 18, wherein above-mentioned at least a chemotherapeutics is by taxol (taxol ^), select in the group that constitutes of Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.

20. in the method for claim 18, wherein above-mentioned at least a IDO inhibitor and at least a chemotherapeutics are used simultaneously.

21. in the method for claim 18, wherein above-mentioned at least a IDO inhibitor and at least a chemotherapeutics are used in order.

22. in the method for claim 21, wherein above-mentioned at least a IDO inhibitor was used before at least a chemotherapeutics.

23. in the method for claim 21, wherein above-mentioned at least a chemotherapeutics was used before at least a IDO inhibitor.

24. in the method for claim 18, wherein above-mentioned chronic viral infection is to select in the group that is made of hepatitis C virus (HCV), Human papilloma virus HPV (HPV), cytomegalovirus (CMV), epstein-Barr virus (EBV), varicella zoster virus, Coxsackie virus and human immunodeficiency virus (HIV).

25. pharmaceutical composition that is used for the treatment of chronic viral infection, wherein above-mentioned composition can accept to carry at least a indoleamine 2 that contains in this medium by certain significant quantity in pharmacy, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutics, wherein above-mentioned at least a IDO inhibitor are to select from the compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups: R wherein _AAnd R _BFrom group H and alkyl, select independently; R wherein _CFrom group H and alkyl, select;

Wherein n is from 1 to 10 integer, R _DIt is molecular formula

The carboline substituting group; With

R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group With The part of the middle ring of selecting, wherein R _EBe alkyl or alkyl-Q, Q represents molecular formula

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

26. in the composition of claim 25, select in the group of wherein above-mentioned at least a chemotherapeutics by taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives formation.

27. in its patient of needs, treat method for cancer for one kind, comprise and side by side or according to priority use at least a indoleamine 2 of certain significant quantity promptly for above-mentioned patient, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutics, wherein above-mentioned at least a IDO inhibitor are to select from one group of compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H;

R ₃By selecting in the following groups: R wherein _AAnd R _BFrom group H and alkyl, select independently; R wherein _CFrom group H and alkyl, select;

Wherein n is from 1 to 10 integer, R _DIt is molecular formula The carboline substituting group; With R wherein _λAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group With The part of the middle ring of selecting, wherein R _EBe alkyl or alkyl-Q, Q represents molecular formula Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II): Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

28. in the method for claim 27, select in the group of above-mentioned at least a chemotherapeutic by taxol (taxol ), Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives formation.

29. in the method for claim 28, above-mentioned at least a chemotherapeutic is a taxol.

30. in the method for claim 27, above-mentioned at least a IDO inhibitor and at least a chemotherapeutics are taken simultaneously.

31. in the method for claim 27, wherein above-mentioned at least a IDO inhibitor and at least a chemotherapeutics are taken in order.

32. in the method for claim 31, wherein above-mentioned at least a IDO inhibitor was taken before at least a chemotherapeutics.

33. in the method for claim 31, wherein above-mentioned at least a chemotherapeutics was taken before at least a IDO inhibitor.

34. in the method for claim 27, wherein above-mentioned cancer is selected from the cancer of following histoorgan: prostate gland, colorectum, pancreas, uterine cervix, stomach, uterine endometrium, brain, liver, bladder, ovary, testis, head, neck, skin (comprising melanoma and rodent cancer), an endo-endothelial layer, white corpuscle (comprising lymphoma and leukemia), esophagus, mammary gland, muscle, reticular tissue, lung (comprising small cell lung cancer and non-small cell carcinoma), suprarenal gland, Tiroidina, kidney or bone; Glioblastoma, mesothelioma, renal cell carcinoma, cancer of the stomach, sarcoma, choriocarcinoma, sebaceous carcinoma and seminoma of testis.

35. pharmaceutical composition that is used for the treatment of cancer, above-mentioned wherein composition contains at least a indoleamine 2 by certain significant quantity in the pharmaceutical acceptable carrier medium, 3-dioxygenase (IDO) inhibitor and at least a chemotherapeutic constitute, and wherein above-mentioned at least a IDO inhibitor is to select from the compound with molecular formula (I) and molecular formula (II):

Molecular formula (I): R wherein ₁Be H or low alkyl group; R ₂Be H; R ₃By selecting in the following groups:

R wherein _AAnd R _BFrom group H and alkyl, select independently;

R wherein _CFrom group H and alkyl, select;

Wherein n is from 1 to 10 integer, R _DIt is molecular formula The carboline substituting group; And R wherein _AAnd R _BFrom group H and alkyl, select independently; Or R ₂And R ₃Combine and represent molecular formula (I) with pyrroles's condensed with from group

With The part of the middle ring of selecting, wherein R _EBe alkyl or alkyl-Q, Q represents molecular formula

Substituting group, the compound of molecular formula (I) is worked as R ₂And R ₃Become beta-carboline derivatives when combining representative (i), work as R ₂And R ₃Combining becomes the brassilexin derivative when representing (ii), work as R in addition ₂And R ₃Combine and become the brassilexin derivative that N-replaces when representing (iii); X, Y and Z can be the same or different, and are from H, halogen, NO ₂And choose in the alkyl; Work as R ₂And R ₃Combine and represent the part of a ring system, Y also may be an isothiocyanate; Condition is that molecular formula (I) does not contain the compound of selecting: 3-(N-methyl-thiohydantoin)-indoles from following material, 3-(N-phenyl-thiohydantoin)-indoles, 3-(N-allyl group-thiohydantoin)-indoles, 5-methyl-brassinin, brassinin, brassilexin, β-Ka Lin, 3-butyl-β-Ka Lin, 6-fluoro-3-methoxycarbonyl-β-Ka Lin, 6-lsothiocyanates-3-methoxycarbonyl-β-Ka Lin, 3-propoxy--β-Ka Lin, 3-carboxyl-β-Ka Lin, 3-carbon propoxy--β-Ka Lin and 3-carbon-uncle-butoxy-β-Ka Lin;

Molecular formula (II):

Wherein X, Y and Z may be the same also may be different, be from by H, halogen, NO ₂And choose in the alkyl; Condition is not contain 3-amino-2-naphthoic acid in the molecular formula (II).

36. in the pharmaceutical composition of claim 14, wherein above-mentioned at least a chemotherapeutics is by taxol (taxol ^), select in the group that constitutes of Platinol, docetaxel, NSC-241240, vincristine(VCR), vinealeucoblastine(VLB), methotrexate, endoxan, CPT-11,5 FU 5 fluorouracil (5-FU), gemcitabine, Emcyt, Carmustine, adriamycin (Zorubicin), etoposide, white arsenic, Mexician scammony and esperamicin derivatives.

37. in the pharmaceutical composition of claim 15, wherein above-mentioned at least a chemotherapeutics is a taxol.

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