WO2019034178A1 - Dna toxicity dimer compound - Google Patents

Abstract

The present invention relates to a dimer compound of DNA toxicity; compared to the benzodiazepine conjugates described in the above technology, the cytotoxic compound according to the present invention displays a much higher in vivo therapeutic index (ratio between the maximum tolerated dose and the minimum effective dose).

Description

DNA toxic dimer compound Field of invention

The present invention relates to novel cytotoxic compounds and medicaments comprising these cytotoxic compounds and cell binding agents. More particularly, the present invention relates to novel benzodiazepine dimer compounds, derivatives thereof, and intermediates thereof which are pharmaceutically acceptable salts, which are useful as medicaments, especially as antiproliferative agents.

Background of the invention

Benzodiazepine derivatives, which have the ability to recognize and bind to specific DNA sequences, are highly potent interchain crosslinkers that react with guanine in the DNA minor groove to form DNA adducts that interfere with The processing of DNA is therefore used as an anti-tumor drug. (Rahman et al. (2009) Jour. Amer. Chem. Soc. 131 (38): 13756-13766; Thurston et al. (1994) Chem. Rev., 94: 433-465; Bose et al. (1992) J. Am. Chem. Soc. 114: 4939-4941; Gregson et al. (2004) Jour. Med. Chem. 47(5): 1161-1174).

The DNA small groove alkylating agent of the 1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indole (CBI) class is a potent cytotoxin (Atwell et al. (1999) J. Med. Chem., 42: 3400), and has been used as an effector unit for a variety of prodrugs designed for cancer therapy. CBI and benzodiazepine derivatives have been linked together via an alkyl chain (Tercel et al. (2003) J. Med. Chem., 46: 2132-2151).

Imidazo[1,2-a]pyridine derivatives are powerful DNA binding units that have been used in the synthesis of anti-tumor antibiotics such as duocarmycins and exhibit very potent cytotoxicity. (Ronald C. Elgersma et al. (2015) Mol. Pharmaceutics. 12: 1813-1835).

The benzodiazepine derivatives disclosed in the prior art are extremely toxic and toxic at very low doses, thus improving the lower toxicity and therapeutically active, high therapeutic window benzodiazepines. Subclass derivatives are very necessary.

Summary of the invention

The present invention aims to provide a cytotoxic benzodiazepine dimer derivative having a good therapeutic window. The newly designed cytotoxic benzodiazepine dimer derivative has two functional groups, chloromethyl (CBI) and imine (PBD) or DNA-binding (DB) and imine (PBD). Chloromethyl is a prodrug structure that forms a three-membered ring structure into the body and can further undergo DNA alkylation. DNA-binding fragments have strong DNA binding. The presence of these two functional groups enhances the crosslinking of the DNA. The design of the present invention surprisingly found that the cytotoxic compound exhibited a much higher in vivo therapeutic index (ratio of the maximum tolerated dose to the lowest effective dose) compared to the benzodiazepine conjugates disclosed in the prior art.

In a general aspect, the present invention provides a cytotoxic drug having the formula (I) or formula (II) or

a pharmaceutically acceptable salt, hydrate or solvate,

among them:

Y is selected from one or more of H, halogen, C _1-4 alkyl, OR ₄ , SR ₄ , NR ₄ R ₅ , SO ₂ R ₄ , CONHR ₄ , CN or COOR ₄ , wherein R ₄ and R ₅ a hydrocarbon group selected from H, C _1-4 , a phenyl group or a substituted phenyl group;

X _{2 is} selected from halogen or OSO ₂ R ₅ , wherein R ₅ may be independently selected from H, C _1-4 hydrocarbyl, phenyl or substituted phenyl;

X _{1 is} selected from O and NR ₆ , wherein R _{6 is} selected from H, C _1-6 alkyl, one or more F substituted C _1-6 alkyl;

R _{1 is} selected from H,P(O) ₃ H ₂ , C(O)NR ₇ R ₈ , wherein R ₇ and R _{8 are} selected from H, C _1-6 alkyl, one or more F substituted C _{1 -6} alkyl, or R ₇ and R ₈ form a five- or six-membered heterocyclyl group;

R ₃ is an optional substituent group.

R _{2 is} selected from H,P(O) ₃ H ₂ , C(O)NR ₇ R ₈ , wherein R ₇ and R _{8 are} selected from H, C _1-6 alkyl, one or more F substituted C _{1 -6} alkyl, or R ₇ and R ₈ form a five- or six-membered heterocyclyl group;

D _{1 is} selected from the following drug modules:

Where the wavy line indicates the site attached to T;

表示单键或双键，条件是当其为双键时，R ₉不存在并且R ₁₀是H，并且当其为单键时，R ₉是H或选自乙酰基、三氟乙酰基、叔丁氧羰基、苄氧羰基和9-芴甲氧羰基的氨基保护部分；R ₁₀选自OH，由O R ₁₁表示的醚、亚硫酸根SO ^3-或OSO ^3-，其中R ₁₁选自具有1-10个碳原子的直链、支链或环状烷基、烯基或炔基； Double line between N and C

Represents a single bond or a double bond, provided that when it is a double bond, R ₉ is absent and R ₁₀ is H, and when it is a single bond, R ₉ is H or is selected from acetyl, trifluoroacetyl, uncle An amino-protecting moiety of butoxycarbonyl, benzyloxycarbonyl and 9-fluorenylmethoxycarbonyl; R _{10 is} selected from OH, an ether represented by O R ₁₁ , sulfite SO ^{3 -} or OSO ^3- , wherein R _{11 is} selected from the group consisting of 1 a linear, branched or cyclic alkyl, alkenyl or alkynyl group of -10 carbon atoms;

The dashed line between C2 and C3 indicates the optional presence of a double bond;

When there is a double bond between C2 and C3,

R _{12 is} selected from C _5-10 aryl optionally substituted by one or more substituents, wherein the substituent is selected from the group consisting of halogen, nitro, cyano, amino, ether, carboxyl, ester, C _{a 1-7} hydrocarbyl group, a C _3-7 heterocyclic group; when a single bond exists between C2 and C3,

其中R ₁₄和R ₁₅独立的选自H、F、C _1-4饱和烷基、C _2-3烯基，所述烷基和烯基被一基团可选取代，所述基团选自C _1-4烷基酰胺基和C _1-4烷基酯，或者，当R ₁₄和R ₁₅之一是H时，另一个选自腈和C _1-4烷基酯； R ₁₂ is

Wherein R ₁₄ and R _{15 are} independently selected from the group consisting of H, F, C _1-4 saturated alkyl, C _2-3 alkenyl, the alkyl and alkenyl optionally substituted by a group selected from the group consisting of a C _1-4 alkyl amide group and a C _1-4 alkyl ester, or, when one of R ₁₄ and R ₁₅ is H, the other is selected from the group consisting of a nitrile and a C _1-4 alkyl ester;

Module D1" may be substituted by one or more R ₁₃ wherein R _{13 is} selected from the group consisting of H, halogen, nitro, cyano, amino, ether, carboxyl, ester, C _1-7 hydrocarbyl, C _3-7 heterocyclyl ;

Preferably, the drug, R3 is selected from the group consisting of H, OH, SH, NH ₂ , N ₃ , NO ₂ , NO, CF ₃ , CN, C(O)NH ₂ , C(O)H, C(O )OH, halogen, R ^h , SR ^h , S(O)R ^h , S(O) ₂ R ^h , S(O)OR ^h , S(O) ₂ OR ^h , OS(O)R ^h , OS( O) ₂ R ^h , OS(O)OR ^h , OS(O) ₂ OR ^h , OR ^h , NHR ^h , N(R ^h )R ⁱ , ⁺ N(R ^h )(R ⁱ )R ^j , P( O)(OR ^h )(OR ⁱ ), OP(O)(OR ^h )(OR ⁱ ), SiR ^h R ⁱ R ^j , C(O)R ^h , C(O)OR ^h , C(O)N (R ^h )R ⁱ , OC(O)R ^h , OC(O)OR ^h , OC(O)N(R ^h )R ⁱ , N(R ^h )C(O)R ⁱ ,N(R ^h ) C(O)OR ⁱ , N(R ^h )C(O)N(R ⁱ )R ^j .

Wherein R ^h , R ⁱ and R ^{j are} independently selected from H and optionally substituted C _1-15 alkyl, C _1-15 heteroalkyl, C _3-15 cycloalkyl, C _1-15 heterocycloalkane a group, a C _5-15 aryl group or a C _1-15 heteroaryl group.

T is selected from a C ₂ -C ₁₂ hydrocarbyl group, Z, (C ₁ -C ₆ alkylene)-Z-(C ₁ -C ₆ alkylene), (C ₁ -C ₆ alkylene)-Z- (C ₁ -C ₆ alkylene)-Z-(C ₁ -C ₆ alkylene), (C ₁ -C ₆ alkenylene)-Z-(C ₁ -C ₆ alkenylene), (C _1- C ₆ alkynylene)-Z-(C ₁ -C ₆ alkynylene);

Wherein Z is independently selected from the group consisting of O, S, NR ₁ , aryl and heteroaryl;

Wherein the alkylene, alkenylene, aryl and heteroaryl groups are independent and optionally F, OH, O(C ₁ -C ₆ alkyl), NH ₂ , NHCH ₃ , N(CH ₃ ) ₂ and C ₁ -C ₆ alkyl substituted wherein the alkyl group is optionally substituted with one or more F;

Preferably, the drug, wherein X ₁ is O, and R ₁ is H;

Preferably, the medicament, wherein D1 is D1 ', the presence of a double bond between the N and C, the presence of a double bond between C2 and C3, and R ₁₂ is methoxy substituted phenyl;

Preferably, the medicament, wherein D1 is D2 ", a double bond exists between the N and C, and R ₁₃ is methoxy;

Preferably, the drug, wherein T is selected from the group consisting of C2-12 alkylene groups;

Preferably, the drug, wherein T is

Preferably, the present invention provides a method of treating a patient in need thereof, comprising administering to the patient a medicament according to any one of the preceding claims, wherein the patient has a tumor, an autoimmune disease or an infectious disease .

The compositions of the invention are useful for inhibiting abnormal cell growth or treating a value-added disease in a mammal, such as a human. The compositions of the invention are useful for treating diseases, such as cancer, in a mammal, such as a human. Wherein the cancer is selected from the group consisting of breast cancer, gastrointestinal cancer, for example, a group consisting of colorectal cancer, pancreatic cancer, cholangiocarcinoma, hepatocellular carcinoma, osteosarcoma, lung cancer, prostate cancer, squamous cell carcinoma, ovarian cancer, testis Cancer, bladder cancer, stomach cancer, head and neck cancer, cervical cancer, kidney cancer, glioma, skin cancer, such as: malignant melanoma, thyroid cancer, leukemia, malignant lymphoma.

Detailed ways

The invention may be more readily understood by reference to the specific embodiments of the invention and the embodiments thereof.

Abbreviations and definitions

Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. When a trade name is used herein, unless otherwise indicated by the context, the trade name includes the product formulation, generic drug, and active pharmaceutical ingredient of the trade name product.

Unless otherwise indicated, the term "aryl" refers to a polyunsaturated, generally aromatic, hydroxyl group which may be monocyclic or fused or covalently attached polycyclic (up to three rings). The term "arylhetero" refers to an aryl (or ring) containing from 1 to 5 heteroatoms selected from N, O or S, wherein the nitrogen and sulfur atoms are optionally oxidized, the nitrogen atom optionally being seasoned Ammonium. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of aryl groups include phenyl, naphthyl and diphenyl, and non-limiting examples of heteroaryl groups include: pyridyl, pyridazinyl, pyrazinyl, pyrimindinyl, Triazinyl, quinolyl, quinoxalinyl, quinazolinyl, porphyrinyl, phthalaziniyl, benzotriazinyl, fluorenyl, benzimidazolyl, benzopyrazole, benzene And triazolyl, benzisoxazolyl, isobenzofuranyl, isodecyl, pyridazinyl, benzotriazinyl, thienopyridyl, thienopyrimidinyl, pyridopyrimidinyl, imidazopyridine , benzothiaxolyl, benzofuranyl, benzothienyl, fluorenyl, quinolyl, isoquinolinyl, isothiazolyl, pyrazolyl, oxazolyl, pteridoyl, imidazolyl , triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, and thienyl. When described as "substituted", the substituents of the above aromatic and heteroaryl ring systems are selected from the following acceptable substituents.

Unless otherwise indicated herein, the substituents of the hydrocarbyl group (including those generally referred to as alkylene, alkenyl, alkynyl and cycloalkyl) may be a plurality of groups selected from the group consisting of -halogen, -OR' , -NR'R", -SR', -SiR'R"R"', -OC(O)R', -C(O)R', -CO2R', -CONR'R", -OC(O )NR'R", -NR"C(O)R', -NR'-C(O)NR"R"', -NR"C(O)2R', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R", -NR'S( O) 2R", -CN and -NO2, the number of substituents is 0 to (2m'+1), where m' is the total number of carbon atoms in the group. R', R" and R"' are independent Hydrogen-substituted, unsubstituted C1-8 alkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted C1-8 alkyl, C1-8 alkoxy or C1-8 thio An alkoxy group, or an unsubstituted aryl-C1-4 alkyl group. When R' and R" are bonded to the same nitrogen atom, they may form a 3-, 4-, 5-, 6- or together with the nitrogen atom. 7-member ring. For example, -NR'R" includes 1-pyrrolidinyl and 4-morpholinyl.

A "derivative" of a compound as used herein refers to a substance having a chemical structure similar to that of a compound but further containing at least one chemical group not present in the compound and/or lacking a chemical group present in at least one compound. The compounds to which the derivatives are compared are referred to as "parent" compounds. Generally, a "derivative" can be produced from a parent compound in one or more chemical reaction steps.

The term "inhibiting" or "inhibiting" means reducing the detectable amount or completely preventing it.

The term "cancer" refers to a physiological condition or disease characterized by dysregulated cell growth. "Tumors" include cancer cells.

The term "autoimmune disease" is a disease or disorder that results from tissue or protein directed against the individual's own body.

The phrase "pharmaceutically acceptable salt" as used herein refers to a pharmaceutically acceptable organic or inorganic salt of the compound. The compound may contain at least one amino or carboxyl group and thus may form an addition salt with the corresponding acid or base. Exemplary salts include, but are not limited to, sulfates, trifluoroacetates, citrates, acetates, oxalates, chlorides, bromides, iodides, nitrates, hydrogen sulfates, phosphates, acids Phosphate, isonicotinic acid, lactate, salicylate, acidic citrate, tartrate, oleate, tannic acid, pantothenate, hydrogen tartrate, ascorbate, salicylate, Formate, orthoformate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, potassium salt, sodium salt, and the like. Additionally, pharmaceutically acceptable salts have more than one dotted atom in the structure. An example in which a plurality of charged atoms are part of a pharmaceutically acceptable salt can have multiple counterexamples. For example, a pharmaceutically acceptable salt has one or more charged atoms and/or one or more counter atoms.

The invention is further illustrated by the following examples, which are intended to illustrate the invention and not to limit the scope of the invention. The test methods which do not specify the specific conditions in the following examples are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer. All percentages, ratios, ratios, or parts are by weight unless otherwise indicated.

All the professions and sciences used herein are used in the same sense as those skilled in the art, unless otherwise defined. In addition, any methods and materials similar or equivalent to those described may be employed in the methods of the invention. The preferred embodiments and materials described herein are for illustrative purposes only.

The general steps employed in the following embodiments of the invention are:

General procedure A

Synthesis of benzodiazepine derivatives

General Step B

Synthesis of benzodiazepine derivatives

General procedure C

Synthesis of CBI derivatives

General Step D

Synthesis of DNA-binding fragments

General procedure E

Synthesis method of CBI-benzodiazepine derivative

General procedure F CBI-benzodiazepine derivative synthesis method two

General procedure G

Binding-benzodiazepine derivative synthesis method

DRAWINGS

Figure 1 is a graph showing the results of the hct116 experiment.

Figure 2 is a graph showing the results of the SKOV3 experiment.

Detailed ways

Example 1 Preparation of 3-(methoxyformyl)bicyclo[1.1.1]pentane-1-carboxylic acid

1.40 g of bicyclo[1.1.1]pentane-1,3-dicarboxylic acid methyl ester was added to a 50 ml three-necked flask, which was dissolved in 20 ml of methanol, and a solution of NaOH (324 mg) in methanol (5 ml) was slowly added dropwise at room temperature. After the completion, the temperature was raised to 80 ° C for 2 h. After completion of the reaction, the mixture was concentrated to dryness. EtOAc (EtOAc) (EtOAc (EtOAc) White solid 925 mg, yield 68%. ¹ H NMR (400 MHz, CDCl 3 ) δ 3.71 (s, 3H), 2.38 (s, 6H); MS m/z 171 (M+1).

Example 2 Preparation of methyl 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylate

In a 500 ml three-necked flask, 6.34 g of 3-(methoxyformyl)bicyclo[1.1.1]pentane-1-carboxylic acid, 30 ml of tetrahydrofuran, 4.89 g of triethylamine were successively added, and the reaction system was cooled to -10 ° C, and slowly added dropwise. 3.87 g of methyl chloroformate was added and reacted at 0 ° C for 1 h, 2.54 g of NaBH 4 was added in portions, and the reaction was continued at 0 ° C until the starting material was completed. The reaction was quenched with EtOAc (EtOAc) (EtOAc (EtOAc) The yield was 66%. ^{1 H NMR (400MHz, CDCl3)} δ3.71 (s, 3H), 2.38 (s, 6H); 3.48 (s, 2H); MS = 157 (M + 1)

Example 3 Methyl 3-((p-toluenesulfonyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate

700 mg of methyl 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylate was added to a 100 ml flask, which was dissolved in 10 ml of dichloromethane, followed by the addition of 1.2 ml of triethylamine and 0.55 g of DMAP, 0 ° C. A dichloromethane solution of p-toluenesulfonyl chloride was added dropwise, and after completion of the addition, the reaction was allowed to proceed until the starting material was completely reacted. The organic layer was washed with EtOAc EtOAc (EtOAc m.

Preparation of Compound 002 of Example 4

Compound 001 (29 g, 67.38 mmol) and 400 ml DCM were added to a 1 L single-mouth bottle, stirred and dissolved, and trichloroisocyanuric acid (23.45 g, 101.07 mmol, TCCA) and tetramethylpiperidine nitrous oxide were added in portions under ice cooling. The material (0.32 g, 2.03 mmol, TEMPO) was added and the mixture was transferred to room temperature overnight. 40 ℃ concentrated under reduced pressure, the residue was dissolved in 500ml of water and 500ml of ethyl acetate, the organic layer was separated, washed with saturated NaHCO ₃ solution, water, and washed once each with saturated brine, dried over anhydrous sodium sulfate, filtered, to Concentration under reduced pressure at 45 ° C to give a yellow solid (34.0 g, m.jjjjjjjjjjjjjjjjjj . Preparation of Compound 003 of Example 5

Compound 002 (15.0 g, 35.01 mmol) and dry DCM 300 ml were added to a 500 ml single-mouth bottle, dissolved and dissolved, N _{2 was} protected and cooled to below -50 ° C, and 2,6-lutidine (16.3 ml, 139.95 was added. After the addition of trifluoromethanesulfonic anhydride (17.7 ml, 105.21 mmol, Tf ₂ O), the mixture was added and reacted at the same temperature for 1.5 h. To the reaction mixture was added ice water 300ml, the organic layer was separated, washed with saturated NaHCO ₃ solution, water, and washed once each with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure at 45 ℃, to give a brown oil 21.5 g, the crude product was mixed with 25 g of silica gel, column chromatography, and a solvent (PE: EA = 5: 1-2:1) to obtain 15.8 g of a yellow oil, yield 80.5%.

Preparation of Compound 004 of Example 6

Compound 003 (13.8 g, 24.62 mmol), 200 ml of EtOH, 200 ml of toluene and 60 ml of water were added to a 1 L three-necked flask, and triethylamine (7.0 ml, 49.85 mmol) and 4-methoxyphenylboronic acid (5.24) were sequentially added with stirring. g, 34.5 mmol), after N ₂ substitution, Pd(PPh ₃ ) _{4 was} added, and after completion, N _{2 was} protected at 30 ° C overnight, and TLC showed that the starting material was completely reacted. The organic layer was separated, and the organic layer was extracted with 100 ml of ethyl acetate. The organic layer was combined and washed with water and saturated brine. The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure at 45 ° C to give 18.2 g of a brown solid. The crude product was mixed with 20 g of silica gel, column chromatography, eluent (PE: EA=4:1-1:1) to obtain yellow The solid was 11.2 g, and the yield was 87.8%.

Preparation of Compound 005 of Example 7

Compound 004 (10.5g, 20.25mmol) and 5% formic acid/methanol solution 250ml were added to a 500ml single-mouth bottle, cooled to below 5 °C in an ice bath, and zinc powder (26.32g, 404.92mmol) was added slowly with stirring. The reaction was heated to 70 ° C under reflux for 8 h, and TLC showed the starting material was completely reacted. After hot filtration, the filter cake was washed with a small amount of methanol, and the filtrate was adjusted to pH 7 with a saturated sodium hydrogen carbonate solution, and then concentrated under reduced pressure at 50 ° C to remove solvent to afford a brownish oil. The mixture was washed with brine, dried over anhydrous sodium sulfate, filtered, evaporated, evaporated, evaporated, evaporated, ), 8.9 g of a yellow solid was obtained in a yield of 96.3%.

Preparation of Compound 006 of Example 8

60% NaH (0.96g, 24.0mmol) and 15ml DMF were added to a 250ml single-mouth bottle, cooled to -10 ° C with stirring, and a mixed solution of compound 005 (8.4 g, 18.4 mmol) and 50 ml of DMF was added dropwise, and the same was added. After stirring at room temperature for 2 h, a mixed solution of 2-(trimethylsilyl)ethoxymethyl chloride (4.0 g, 24.0 mmol) and 15 ml of DMF was added dropwise to the reaction mixture, and the reaction was carried out at -5 ° C overnight. TLC showed the starting material to react completely. The reaction mixture was poured into 300 ml of ice water, and extracted with 200 ml of DCM. The organic layer was washed successively with saturated NaHCO ₃ solution, water and brine, dried over anhydrous sodium sulfate, filtered and evaporated 10.5 g of an oily product, the crude product was mixed with 15 g of silica gel, column chromatography, and the solvent (DCM: MeOH = 600: 1-400:1) to give 6.1 g of a yellow solid.

Preparation of Compound 007 of Example 9

Compound 006 (2.0 g, 3.41 mmol) and 30 ml of MeOH were added to a 250 ml one-necked flask. After stirring and stirring, HCO ₂ NH ₄ (2.15 g, 34.10 mmol) and 5% Pd/BaSO ₄ (0.28 g) were added in that order. After reacting for 4 h at room temperature, TLC showed that the starting material was completely reacted. The reaction solution was filtered through celite, and the filter cake was washed with a small amount of DCM. The filtrate was concentrated and concentrated under reduced pressure at 45 ° C. The residue was dissolved in 100 ml of DCM. Filtration and concentration under reduced pressure at 40 ° C to give 1.59 g of pale yellow solid. The crude material was mixed with 2.0 g of silica gel, column chromatography, eluent (PE: EA = 3: 1-1:1) to give a yellow solid 1.45 g The yield was 85.8%.

Synthesis of Example 10 Intermediate 008

Add methyl 3-((p-toluenesulfonyloxy)methyl)bicyclo[1.1.1]pentane-1-carboxylate (0.92 g, 1.85 mmol), Intermediate 007 (0.61 g, 1.96 mmol) and 20 ml of DMF were stirred and dissolved, and then cesium carbonate (1.21 g, 3.71 mmol) was added. After the addition, the temperature was raised to 80 ° C for 4 h under nitrogen atmosphere, and TLC showed that the starting material was completely reacted. 60 ml of water was added to the reaction mixture, and the mixture was extracted twice with 30 ml of EA. The organic layer was combined and washed with water and brine, and the organic layer was dried over anhydrous sodium sulfate. 1.4 g of a yellow solid was obtained, and the crude product was mixed with 2.0 g of silica gel, column chromatography, and a solvent (PE: EA = 4: 1-2:1) to obtain a pale yellow solid (1.08 g, yield: 92.3%).

Example 11 Synthesis of Intermediate 009

Intermediate 008 (0.9 g, 1.42 mmol), 15 ml of THF and 5 ml of MeOH were added to a 100 ml one-necked flask, and 1N LiOH (3.0 ml, 3.0 mmol) was added thereto with stirring, and the reaction was carried out under nitrogen atmosphere at room temperature overnight. The reaction is complete. Concentrated under reduced pressure at 45 ° C, the residue was dissolved in 100 ml of DCM, and the mixture was adjusted to pH 4-5 with 1N HCl solution. The organic layer was separated and washed once with water and saturated brine. The sodium was dried, filtered, and concentrated under reduced pressure <RTI ID=0.0></RTI> to <RTI ID=0.0></RTI> </RTI> <RTI ID=0.0> The white solid was 0.75 g, and the yield was 85.2%.

Example 12 Synthesis of Intermediate 010

Intermediate 009 (650 mg, 1.047 mmol) and 15 ml of DMF were added to a 100 ml vial, and 1-ethyl-(3-dimethylaminopropyl)carbonyldiimide hydrochloride (610 mg, 3.182mmol) and DMAP (385mg, 3.151mmol), N 2 protection stirred at room temperature 1h, and then added CBI-10 (470mg, 1.255mmol) , N 2 protection reaction at room temperature overnight, TLC showed the starting material reaction is substantially complete. 40 ml of water was added to the reaction solution, and the mixture was extracted twice with 30 ml of DCM. The organic layer was combined, washed with water and brine, and then evaporated. 1.8 g of a brownish-yellow oil was obtained, and the crude product was mixed with 2.5 g of silica gel, column chromatography, and a solvent (PE: EA = 4: 1-1:1) to give 0.71 g of a pale yellow solid, yield 72.08%.

Example 13 Synthesis of Intermediate 011

Intermediate 010 (0.6 g, 0.638 mmol), 10 ml THF and 20 ml MeOH were added to a 100 ml one-necked flask, stirred and dissolved, and HCO ₂ NH ₄ (0.41 g, 6.502 mmol) and 5% Pd/BaSO ₄ (0.085) were sequentially added at room temperature. g), after adding, reacting at room temperature for 5 h, TLC showed that the starting material was completely reacted. The reaction solution was filtered through celite, and filtered, and then filtered, and then filtered and evaporated. EA: PE = 1:1-2:1) gave 0.3 g of pale yellow solid, yield 55.35%.

Synthesis of Compound 012 of Example 14

Add intermediate 011 (0.13g, 0.153mmol) to 50ml three-necked flask, vacuum with oil pump, switch N ₂ times, inject 5ml of freshly re-distilled THF, stir and dissolve and cool down to -78 °C, then 1N Li ^. BHEt ₃ (1.2 ml, 1.20 mmol) was slowly added dropwise, and the reaction was continued for 2 h at the same temperature. TLC showed that the starting material was completely reacted. After adding 20 ml of DCM, the reaction solution was transferred to an ice bath, and 20 ml of water was added dropwise to quench the reaction. The organic layer was separated, and the aqueous layer was extracted once with 10 ml of DCM. The organic layer was combined and washed once with water and saturated brine. Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure at 40 ° C to give 152 mg of pale yellow solid. Immediately add 3 ml of DCM, 6 ml of CH ₃ OH and 1 ml of H ₂ O, stir and dissolve, then add silica gel until the mixture is viscous. After sealing, it was stirred at room temperature overnight. After adding 15 ml of DCM and 5 ml of CH ₃ OH, it was filtered, and the filter cake was washed with a small amount of DCM. The filtrate was concentrated and concentrated under reduced pressure at 45 ° C to give 0.12 g of pale yellow solid. The compound (DCM: MeOH = 60:1 - 30:1) gave 25 mg of pale yellow solid.

¹ H-NMR (CDCl ₃ ): δ=8.46 (s, 1H), 8.23 (d, 1H), 7.88 (d, 1H), 7.58 (s, 1H), 7.29-7.39 (m, 6H), 6.86- 6.92 (m, 3H), 4.02-4.61 (m, 5H), 4.00 (s, 3H), 3.83 (s, 3H), 3.49 (s, 2H), 2.76 (s, 3H), 1.62 (m, 8H) .

Example 15: 5-((7-(Methoxy)-2-(4-methoxyphenyl)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)) Methyl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentanoic acid Ester (Compound 013)

0.1 g of 7-(methoxy)-8-hydroxy-2-(4-methoxyphenyl)-10-((2-(trimethylsilyl)ethoxy)methyl)- was added to a 25 ml flask. 1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dioxo (compound 007) and 4 ml DMF Under stirring, 42 mg of K2CO3 and 0.1 ml of ethyl 5-bromo-valerate were sequentially added, and the mixture was reacted at room temperature until the starting material was completed. After adding 15 ml of water, the mixture was extracted with ethyl acetate. It was used directly in the next reaction without purification.

Example 16: Compound 014: 5-((7-(Methoxy)-2-(4-methoxyphenyl)-5,11-dioxo-10-((2-(trimethylsilyl)) Oxy)methyl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy) Valeric acid

0.2 g of 5-((7-(methoxy))-2-(4-methoxyphenyl)-5,11-dioxo-10-((2-(trimethylsilyl))) was added to a 50 ml flask. Oxy)methyl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy) Ethyl valerate (Compound 013) was dissolved in THF / MeOH (V: V = 3: 1), and 1N LiOH was added with stirring, and the mixture was reacted at room temperature until the starting material was completed. The solvent was concentrated under reduced pressure at 40 ° C. The residue was dissolved in dichloromethane. The residue was taken to dichloromethane to pH 5 to 6 and then the organic layer was separated. The mixture was washed with brine, dried over anhydrous sodium sulfate. Column chromatography purification (DCM: MeOH = 30:1)ield

Preparation of Compound 015 of Example 17

20 mg of 5-((7-(methoxy))-2-(4-methoxyphenyl)-5,11-dioxo-10-((2-(trimethylsilyl))ethoxy group was added to a 25 ml flask. )methyl)-5,10,11,11a-tetrahydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-8-yl)oxy)pentanoic acid (Compound 014), after dissolving it with 1 ml of DMF, adding 10 mg of EDCI and 5 mg of DMAP, stirring at room temperature for 1 h, followed by the addition of 14 mg of 5-(benzyloxy)-1-(chloromethyl)-9-methyl-2. 3-Dihydro-1H-benzo[e]indole, reacted at room temperature until the starting material is completely reflected. The reaction mixture was poured with water, and the mixture was evaporated. Column chromatography purification (petroleum ether: ethyl acetate = 3:1) gave a yellow solid (yield: 30%).

Preparation of Compound 016 of Example 18

10 mg of compound 015 and MeOH/THF (V: V = 1:1) were added to a 25 ml flask, and 7 mg of ammonium formate and 1.4 mg of 5% palladium sulphate were successively added under stirring, and the reaction mixture was completely reacted at room temperature. The reaction solution was filtered through celite, and then filtered and evaporated.

Preparation of Compound 017 of Example 19

50 mg of compound 016 was added to a 50 ml flask, which was dissolved in 2 ml of THF and cooled to -78 ° C, and a solution of lithium triethylborohydride in tetrahydrofuran was slowly added dropwise thereto, and the reaction was completed at -78 ° C until the starting material was completely reacted. The reaction solution was slowly added dropwise with H ₂ O. The pH was adjusted to 5 to 6 with dilute hydrochloric acid, and then extracted three times with dichloromethane. The organic layer was combined and dried and concentrated to give a yellow solid. The obtained solid was dissolved in a mixed solvent of methanol/dichloromethane (4 ml + 1.5 ml), and 0.5 ml of water was added thereto with stirring, followed by the addition of 3 g of silica gel (300-400 mesh), and the reaction was stirred at room temperature. After the completion of the reaction, the reaction mixture was filtered, and then filtered and evaporated. The obtained solid was separated and purified by preparative liquid phase to give Compound 009 12.5 mg.

Example 20 Compound 018: (S)-1-(4-(Benzyloxy)-5-methoxy-2-nitrobenzoyl)-6-methoxyindoline-2-carboxylic acid Preparation of ester

110 mg of 4-(benzyloxy)-5-methoxy-2-nitrobenzoic acid was dissolved in 3 ml of dichloromethane, 62 μl of oxalyl chloride was added at 0 ° C, and the mixture was allowed to react at room temperature for 3 hours, and the reaction solution was concentrated. Light yellow solid. 50 mg of (S)-6-methoxyindoline-2-carboxylic acid methyl ester was dissolved in 5 ml of dichloromethane, 0.1 ml of triethylamine was added at 0 ° C, and the obtained solid solution of dichloromethane was slowly added dropwise. , reacted at 0 ° C for 2 h. The reaction mixture was adjusted to pH 4 to 5 with dilute hydrochloric acid, and extracted twice with ethyl acetate. The obtained solid was purified by column chromatography (yield: ethyl ether: ethyl acetate = 20:1).

Example 21(S)-9-(Benzyloxy)-3,8-dimethoxy-12a,13-dihydro-6H-benzo[5,6][1,4]diazepine [1,2-a]吲哚-6,12(11H)-dione (compound 019)

50 mg of (S)-1-(4-(benzyloxy)-5-methoxy-2-nitrobenzoyl)-6-methoxyindoline-2-carboxylic acid methyl ester was added to a 25 ml flask. It was dissolved in 5 ml of methanol, and 96.4 mg of stannous chloride was added thereto with stirring, and the temperature was raised to 65 ° C until the starting material was completed. The reaction was cooled, washed with saturated NaHCO ₃ PH adjusted to 7 ~ 8, and then filtered through Celite, the filtrate was concentrated diluted with water, and extracted twice with ethyl acetate, the organic layers were combined, dried and concentrated and saturated brine, A pale yellow solid was obtained (yield: 63%).

Example 22(S)-9-(Benzyloxy)-3,8-dimethoxy-11-((2-(trimethylsilyl)ethoxy)methyl)-12a,13-dihydro- 6H-Benzo[5,6][1,4]diazepine[1,2-a]indole-6,12(11H)-dione (compound 020).

26 mg of sodium hydride was suspended in anhydrous DMF, and 215 mg of (S)-9-(benzyloxy)-3,8-dimethoxy-12a,13-dihydro-6H-benzo[ 5,6][1,4]Diazepolo[1,2-a]indole-6,12(11H)-dione in DMF, stirred for 30 min, then slowly added 108 mg of SEMCl The DMF solution is added to the same temperature to complete the reaction. The reaction mixture was poured into ice water and extracted with EtOAc EtOAc (EtOAc)EtOAc. ), 195 mg of oil was obtained in a yield of 72%.

Example 23(S)-9-(hydroxy)-3,8-dimethoxy-11-((2-(trimethylsilyl)ethoxy)methyl)-12a,13-dihydro-6H- Benzo[5,6][1,4]diazepine[1,2-a]indole-6,12(11H)-dione (compound 021)

190 mg of (S)-9-(benzyloxy)-3,8-dimethoxy-11-((2-(trimethylsilyl)ethoxy)methyl)-12a, 13-di was added to a 25 ml flask. Hydrogen-6H-benzo[5,6][1,4]diazepine[1,2-a]indole-6,12(11H)-dione, dissolved in 10 ml of methanol, at room temperature 214 mg of ammonium formate, 95 mg of 5% palladium sulphate were added in sequence, and the reaction was completed under nitrogen to complete the starting material. The reaction mixture was filtered over EtOAc (EtOAc)EtOAc. .

Example 24 3-(((8-(Benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl)ethoxy)methyl)) -11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]indol-9-yl)oxy)methyl) Bicyclo [1.1.1] methyl pentane-1-carboxylate (compound 022)

30 mg of (S)-9-(hydroxy)-3,8-dimethoxy-11-((2-(trimethylsilyl)ethoxy)methyl)-12a,13-dihydro- 6H-benzo[5,6][1,4]diazepine[1,2-a]indole-6,12(11H)-dione and 2 ml of DMF, and added Cs ₂ CO ₃ with stirring. The reaction was heated to 80 ° C to complete the reaction. After adding 20 ml of water, the mixture was extracted with ethyl acetate. It was used directly in the next reaction without purification.

Example 25 3-(((8-(Benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl)ethoxy)methyl)) -11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]indol-9-yl)oxy)methyl) Bicyclo[1.1.1]pentane-1-carboxylic acid (compound 023)

36 mg of 3-(((8-(benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl))ethoxy)) was added to a 25 ml flask. Methyl)-11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]fluoren-9-yl)oxy) Methyl)dicyclo[1.1.1]pentane-1-carboxylic acid methyl ester was dissolved in THF/MeOH (V:V = 3:1), and 1N LiOH was added with stirring. The solvent was concentrated under reduced pressure at 40 ° C, the residue was dissolved in dichloromethane, and then the mixture was evaporated to m~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ The organic layer was washed with EtOAc EtOAc EtOAc.

Preparation of Compound 024 of Example 26

20 mg of 3-((8-(benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl))ethoxy)) was added to a 25 ml flask. Base)-11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]fluoren-9-yl)oxy) Bicyclo[1.1.1]pentane-1-carboxylic acid, dissolved in 2 ml of DMF, added with 20 mg of EDCI and 3 mg of DMAP, stirred at room temperature for 1 h, followed by the addition of 17 mg of 5-(benzyloxy)-1-(chloro Methyl)-9-methyl-2,3-dihydro-1H-benzo[e]indole, reacted at room temperature until the starting material is completely reflected. The reaction mixture was combined with EtOAc. The yield was 34%.

Preparation of Compound 025 of Example 27

10 mg of compound 024 and MeOH/THF (V: V = 1:1) were added to a 25 ml flask, and 7 mg of ammonium formate and 1.4 mg of 5% palladium sulphate were sequentially added under stirring, and the reaction mixture was completely reacted at room temperature. The reaction solution was filtered through celite, and then filtered and evaporated.

Preparation of Compound 026 of Example 28

20 mg of the compound 025 was added to a 50 ml flask, which was dissolved in 2 ml of THF and cooled to -78 ° C, and a solution of lithium triethylborohydride in tetrahydrofuran was slowly added dropwise thereto, and the reaction was completed at -78 ° C until the starting material was completely reacted. The reaction solution was slowly added dropwise with H 2 O. The pH was adjusted to 5 to 6 by dilute hydrochloric acid, and then extracted with dichloromethane for 3 times. The organic layer was combined and dried and concentrated to give a yellow solid. The obtained solid was dissolved in a mixed solvent of methanol/dichloromethane (4 ml + 1.5 ml), and 0.5 ml of water was added thereto with stirring, followed by the addition of 3 g of silica gel (300-400 mesh), and the reaction was stirred at room temperature. After the completion of the reaction, the reaction mixture was filtered, and then filtered and evaporated. The obtained solid was separated and purified by preparative liquid phase to give compound 026 8 mg.

Example 29 5-((8-(Benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl)ethoxy)methyl)-) 11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]indol-9-yl)oxy)pentanoic acid ethyl ester Compound 027

500 mg of (S)-9-(hydroxy)-3,8-dimethoxy-11-((2-(trimethylsilyl)ethoxy)methyl)-12a,13-dihydro- 6H-benzo[5,6][1,4]diazepine[1,2-a]indole-6,12(11H)-dione and 20 ml of DMF, and 200 mg of K2CO3 and 0.5 were added in sequence with stirring. Methyl 5-bromo-pentanoate, added to the reaction at room temperature until the starting material is complete. After adding 100 ml of water, the mixture was extracted with ethyl acetate. It was used directly in the next reaction without purification.

Example 30 5-((8-(Benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl)ethoxy)methyl)-) 11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]fluoren-9-yl)oxy)pentanoic acid (compound) 028)

0.5 g of 5-((8-(benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl))ethoxy) was added to a 50 ml flask. Methyl)-11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]fluoren-9-yl)oxy) Ethyl valerate was dissolved in THF/MeOH (V: V = 3:1), and 1N LiOH was added with stirring, and the reaction was carried out at room temperature until the starting material was completed. The solvent was concentrated under reduced pressure at 40 ° C, the residue was dissolved in dichloromethane, and then the mixture was evaporated to m~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ The organic layer was washed with EtOAc EtOAc EtOAc.

Preparation of Compound 029 of Example 31

200 mg of 5-((8-(benzyloxy)-3-methoxy-6,12-dioxo-11-((2-(trimethylsilyl)ethoxy)methyl) was added to a 250 ml flask. )-11,12,12a,13-tetrahydro-6H-benzo[5,6][1,4]diazepine[1,2-a]fluoren-9-yl)oxy)pentanoic acid After dissolving it in 10 ml of DMF, 100 mg of EDCI and 50 mg of DMAP were added, and stirred at room temperature for 1 h, followed by the addition of 150 mg of 5-(benzyloxy)-1-(chloromethyl)-9-methyl-2,3-dihydro- 1H-benzo[e]pyrene, reacted at room temperature until the starting material is completely reflected. The reaction mixture was added with water and EtOAc (EtOAc m. .

Preparation of Compound 030 of Example 32

15 mg of the compound 013 and MeOH/THF (V: V = 1:1) were added to a 25 ml flask, and 10 mg of ammonium formate and 2 mg of 5% palladium sulphate were sequentially added thereto with stirring, and the reaction was carried out at room temperature until the starting reaction was completed. The reaction solution was filtered through celite, and then filtered, and then filtered and evaporated.

Preparation of Compound 031 of Example 33

30 mg of compound 030 was added to a 25 ml flask, which was dissolved in 2 ml of THF and cooled to -78 ° C. A solution of lithium triethylborohydride in tetrahydrofuran was added dropwise, and the reaction was completed at -78 ° C until the starting material was completely reacted. The reaction solution was slowly added dropwise with H ₂ O. The pH was adjusted to 5-6 with dilute hydrochloric acid and extracted three times with dichloromethane. The organic layer was combined and dried and concentrated to give a yellow solid. The obtained solid was dissolved in a mixed solvent of methanol/dichloromethane (4 ml + 1.5 ml), and 0.5 ml of water was added thereto with stirring, followed by the addition of 3 g of silica gel (300-400 mesh), and the reaction was stirred at room temperature. After the completion of the reaction, the reaction mixture was filtered, and then filtered and evaporated. The obtained solid was separated and purified by preparative liquid phase to give Compound 031 8 mg.

Example 34 Synthesis of Intermediate 032

4-Benzyloxy-5-methoxy-2-nitrobenzoic acid (5.73 g, 18.88 mmol) and 50 ml of dichloromethane were added to a 150 ml vial, 0.1 ml of DMF was added with stirring, and the mixture was cooled in an ice bath. Add oxalyl chloride (3.2 ml, 37.8 mmol), add the reaction, and move the reaction solution to room temperature for 4 h. TLC showed that the starting material was completely reacted. Concentrate under reduced pressure at 45 ° C to remove dichloromethane and the residue was dissolved in 30 mL THF.

An additional 150 ml vial was added, and (S)-6-nitro-porphyrin-2-carboxylic acid methyl ester (2.8 g, 12.6 mmol) and 30 ml of THF were added, and triethylamine (5.3 ml, 38.2 mmol) was added with stirring. Then, the THF solution of the above-prepared acid chloride was added dropwise under ice-cooling, and the mixture was heated to room temperature overnight. The organic layer was washed with 1N HCl, water and brine, and then dried over anhydrous sodium sulfate, filtered and dried at 40 ° C. The mixture was concentrated under pressure to give 9.5 g of a brown solid. The crude material was applied to a mixture of 11.0 g of silica gel, column chromatography, eluent (PE: EA = 5: 1-1:1) to give 6.1 g of a yellow solid.

Example 35 Synthesis of Intermediate 033

Add intermediate 32 (4.9 g, 9.66 mmol) and 5% formic acid/methanol solution 150 ml to a 250 ml one-necked flask, cool to below 5 ° C in an ice bath, and slowly add zinc powder (12.56 g, 193.23 mmol) with stirring. The reaction was heated to 70 ° C under nitrogen for reflux overnight, and TLC showed that the starting material was completely reacted. Filtration with hot water, the filter cake was washed with a small amount of methanol, and the filtrate was adjusted to pH 8-9 with saturated sodium hydrogen carbonate solution, then concentrated under reduced pressure at 50 ° C to remove solvent to afford 6.2 g of brownish oil. The mixture was combined with silica gel, EtOAcjjjjjjjj

Example 36 Synthesis of Intermediate 034

Add 60% NaH (0.38g, 9.5mmol) and 10ml DMF to a 250ml single-mouth bottle, stir

After cooling to -10 ° C, a mixed solution of Intermediate 2 (3.0 g, 7.23 mmol) and 50 ml of DMF was added dropwise, and stirred at the same temperature for 1 h, then 2-(trimethylsilyl)ethoxymethyl chloride (1.67 ml) A mixed solution of 9.41 mmol) and 10 ml of DMF was added dropwise to the reaction solution, and after completion, the reaction was carried out at the same temperature for 5 hours, and TLC showed that the starting material was completely reacted. The reaction solution was poured into 250 ml of water and extracted twice with 200 ml of DCM. The organic layer was washed with saturated NaHCO ₃ solution, water and brine, and then organic layer was evaporated. Concentration under reduced pressure afforded EtOAc (EtOAc: EtOAc. .

Example 37 Synthesis of Intermediate 035

Add intermediate 3 (2.5 g, 4.59 mmol), 30 ml THF and 30 ml MeOH to a 150 ml one-necked flask, stir and dissolve, and add HCO ₂ NH ₄ (2.89 g, 45.87 mmol) and 10% Pd/C (1.25 g) at room temperature. After the addition, the reaction was carried out for 3 h at room temperature, and TLC showed that the starting material was completely reacted. The reaction solution was filtered through celite, and filtered, and filtered, and then filtered, and then filtered, and then evaporated. MeOH = 60:1-30:1) gave 1.63 g of a yellow solid, yield 78.3%.

Example 38 Synthesis of Intermediate 036

Add 5-tert-butoxyamidopentanyl p-toluenesulfonate (900 mg, 2.52 mmol) and 10 ml of DCM to a 100 ml vial, stir to dissolve and cool to below 5 ° C, add 25 ml of 3 mol / L HCl / EA solution, add After completion, the temperature was raised to room temperature under nitrogen atmosphere for overnight reaction, and TLC showed that the starting material was completely reacted. Concentration under reduced pressure at 45 ° C gave 670 mg of white oil.

Synthesis of Intermediate 037

BDN-001 (430 mg, 1.11 mmol) and 40 ml of DMF were added to a 150 ml single-mouth bottle, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (640 mg, 3.33 mmol) and DMAP (410 mg, 3.35 mmol) were stirred at room temperature for 2 h, then intermediate 5 (660 mg, 2.24 mmol). 100 ml of water was added to the reaction solution, and the mixture was extracted twice with 100 ml of DCM. The organic layer was washed with water and brine, and then the organic layer was dried over anhydrous sodium sulfate, filtered and evaporated 1.5 g of a yellow oil, the crude product was applied to a mixture of EtOAc (yield: EtOAc, EtOAc (EtOAc)

Example 39 Synthesis of Intermediate 038

Intermediate 4 (0.29 g, 0.637 mmol) and 10 ml of DMF were added to a 100 ml vial, stirred and dissolved, then cesium carbonate (0.52 g, 1.596 mmol) was added, stirred for 30 min, then intermediate 6 (0.4 g, 0.638 mmol) was added. After the addition, the temperature was raised to 80 ° C under nitrogen for 6 h, and TLC showed that the starting material was completely reacted. 40 ml of water was added to the reaction solution, and the mixture was extracted twice with 50 ml of DCM. The organic layer was washed with water and brine, and the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and evaporated The yellow oil was 1.1 g, and the crude material was crystallised eluted with EtOAc (EtOAc: EtOAc (EtOAc)

Synthesis of Example 40 Intermediate 039

Intermediate 7 (0.3 g, 0.33 mmol), 8 ml THF and 8 ml MeOH were added to a 50 ml one-necked flask, stirred and dissolved, and HCO ₂ NH ₄ (0.21 g, 3.33 mmol) and 5% Pd/BaSO ₄ (0.06) were sequentially added at room temperature. g), after adding, reacting at room temperature for 3 h, TLC showed that the starting material was completely reacted. The reaction solution was filtered through celite, and filtered, and filtered, and then filtered, and then filtered and evaporated. : MeOH = 30: 1-10: 1) gave 0.18 g of pale yellow solid.

Synthesis of Compound 040 of Example 41

Add intermediate 8 (80 mg, 0.098 mmol) to a 50 ml three-necked flask, vacuum with an oil pump, switch N ₂ times, inject 4 ml of freshly re-distilled THF, stir and dissolve, and cool to below -78 ° C, then slowly 1N Li ^. BHEt ₃ (0.98 ml, 0.98 mmol) was added dropwise, and the reaction was continued for 3 h at the same temperature. TLC showed that the starting material was completely reacted. After adding 15 ml of DCM, the reaction solution was transferred to an ice bath, and 20 ml of water was added dropwise to quench the reaction. The organic layer was separated, and the aqueous layer was extracted once with 10 ml of DCM. The organic layers were combined and washed once with water and saturated brine. Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure at 40 ° C to give a yellow solid 105 mg. Immediately add 3 ml of DCM, 6 ml of CH ₃ OH and 1 ml of H ₂ O, stir and dissolve, then add silica gel until the mixture is viscous, sealed. It was then stirred at room temperature overnight. After adding 10 ml of DCM and 3 ml of CH ₃ OH, the mixture was filtered, and the filter cake was washed with a small amount of DCM. The filtrate was combined and concentrated under reduced pressure at 45 ° C to give a pale yellow solid (85 mg). DCM: MeOH = 40:1 - 15:1) gave 27 mg as pale yellow solid.

Synthesis of Compound 041 of Example 42

4 g of the raw material BDN-001 was dispersed in 50 ml of anhydrous dichloromethane (new system), 2.5 ml of oxalyl chloride was added dropwise under ice bath (0 ± 5 ° C), and 0.5 ml of DMF was added dropwise to form bubbles. The mixture was slowly warmed to room temperature and stirred for 2 h. (TLC EtOAc: EtOAc: EtOAc:EtOAc:

The fresh acid chloride was mixed with 50 ml of absolute ethanol and 4 ml of triethylamine in an ice bath (0 ± 5 ° C), and stirred for 2 h in an ice bath (TLC monitoring: starting material reaction completely DCM / MeOH, 10:1). Dilute hydrochloric acid was adjusted to pH neutrality, filtered, and the crude product was recrystallized from absolute ethanol, and concentrated under vacuum at 55 ± 5 ° C (≤ -0.09 MPa) to obtain a yellow solid, 3.1 g, yield 72% (LC-MS confirmed to be the target product)

Synthesis of Compound 042 of Example 43

5 g of the starting material 041 was dissolved in 45 ml of fresh 5-amino-1-pentanol, 100 ml of ethanol was added, and the temperature was slowly raised to 78 ± 5 ° C, and stirred for 4 hours.

(TLC monitoring: raw material reaction complete DCM / MeOH 20:1 expansion 4 times) Add appropriate amount of water to the reaction solution, adjust the pH to 6 with 6M hydrochloric acid in ice bath (0 ± 5 ° C), extract 3 times with ethyl acetate, organic The phase is further washed once with 5% hydrochloric acid, once with saturated NaCl, dried under reduced pressure of 40 ± 5 ° C, concentrated under vacuum at 55 ± 5 ° C (≤ -0.09 MPa) to obtain a yellow solid, 4.1 g, yield 77%, no need Purification was used directly in the next reaction.

Preparation of Example 44 Compound 043

The glass bottle is dried, N2 is replaced, 5 g of the raw material 042 is dispersed in 150 ml of anhydrous dichloromethane (new system), 4 ml of DMF is added to promote dissolution, and PBr ₃ 4.3 g is slowly added dropwise in an ice bath (0±5 ° C). Gradually dissolve. Maintain the temperature for 2 h, if the raw material is left, add PBr ₃ until the reaction is complete (TLC monitoring: the reaction of the raw material is complete DCM / MeOH 20:1 2 times). Adjust the pH with a saturated NaHCO3 solution in an ice bath (0 ± 5 ° C). 7. A large amount of solid was produced, filtered, and purified by column chromatography (DCM / MeOH 100:1) to yield 3 g of product, yield 54% (LC-MS confirmed as the target product).

Preparation of Example 45 Compound 044

The glass bottle is dried, and 1 g of the raw material is 7-(methoxy)-8-hydroxy-2-(4-methoxyphenyl)-10-((2-(trimethylsilyl)ethoxy)methyl) -1,11a-dihydro-5H-benzo[e]pyrrolo[1,2-a][1,4]diazepine-5,11(10H)-dioxo (compound 007) and Dissolve with ultra dry DMF, add 0.79 g of cesium carbonate for 30 min, add 1.5 g of starting compound 043 and 0.08 g KI 50 ± 5 ° C and stir overnight (TLC monitoring: raw material 007 reaction complete DCM / MeOH 30:1), cool to room temperature, add Appropriate amount of water, ethyl acetate extraction 4 times, saturated NaCl washed once, concentrated to give a crude product. Column chromatography purification (DCM / MeOH 100: 1 - 50: 1) gave 1.2 g of product (yield: 63%, (LC-MS)

Preparation of Example 46 Compound 045

1 g of the starting material 044 was dispersed in 50 ml of methanol and dissolved by heating. After cooling to room temperature, 1.32 g of ammonium formate, 0.25 g of 5% Pd-BaSO _{4 was added} , and the pH was adjusted to 7,35 ± 5 ° C with an appropriate amount of 5% formic acid/methanol, and the next day. Add 0.25 g of 5% Pd-BaSO _{4 and} continue to stir for 24 h (TLC monitoring: about 10% unreacted DCM/MeOH in the starting material, 2 times in 20:1). Cool to room temperature, filter through Celite, filter cake with methanol: The dichloromethane was washed 10:1, and the filtrate was concentrated under reduced pressure of 40. Purification by column chromatography (DCM / MeOH 40: 1 - 20:1) afforded 0.5 g of product, material recovered, yield 56%, (LC-MS confirmed as the target product).

Preparation of Compound 046 of Example 47

Dry the glass bottle, under absolute N2 protection, dissolve 30mg of raw material 045 into absolute anhydrous tetrahydrofuran (new system), cool down to -78 °C, slowly add 0.07ml of lithium triethylborohydride solution, stir for 1h-2h, depending on In the case, a solution of lithium triethylborohydride was added. (1M newly opened 25ml specification package), (TLC, HPLC tracking monitoring: raw material reaction complete DCM / MeOH 10:1 expansion 2 times) maintain the temperature by adding appropriate amount of water quenching, add 5% citric acid solution to adjust the pH to 7, acetic acid B The ester was extracted 3 times, washed once with saturated NaCl, and concentrated at 40 ± 5 ° C to give 25 mg of crude material. Semi-preparation gave 10 mg of product BL-BDN-007 in a yield of 35%. (LC-MS confirmed the target product).

Example 56 Toxin Cell Activity Assay

Compound Hct116 SKOV-3 026 (PD-02) 0.36nm 1.04nm 040 (PD-01) 209.8m >2000 046 (PD-03) 253.0nm >2000

Claims (9)

A cytotoxic drug having the formula (I) or formula (II) or a pharmaceutically acceptable salt, hydrate or solvate thereof,

among them: Y is selected from one or more of H, halogen, C _1-4 alkyl, OR ₄ , SR ₄ , NR ₄ R ₅ , SO ₂ R ₄ , CONHR ₄ , CN or COOR ₄ , wherein R ₄ and R ₅ a hydrocarbon group selected from H, C _1-4 , a phenyl group or a substituted phenyl group; X _{2 is} selected from halogen or OSO ₂ R ₅ , wherein R ₅ may be independently selected from H, C _1-4 hydrocarbyl, phenyl or substituted phenyl; X _{1 is} selected from O and NR ₆ , wherein R _{6 is} selected from H, C _1-6 alkyl, one or more F substituted C _1-6 alkyl; R _{1 is} selected from H,P(O) ₃ H ₂ , C(O)NR ₇ R ₈ , wherein R ₇ and R _{8 are} selected from H, C _1-6 alkyl, one or more F substituted C _{1 -6} alkyl, or R ₇ and R ₈ form a five- or six-membered heterocyclyl group; R ₃ is an optional substituent group; R _{2 is} selected from H,P(O) ₃ H ₂ , C(O)NR ₇ R ₈ , wherein R ₇ and R _{8 are} selected from H, C _1-6 alkyl, one or more F substituted C _{1 -6} alkyl, or R ₇ and R ₈ form a five- or six-membered heterocyclyl group; D _{1 is} selected from the following drug modules:

Where the wavy line indicates the site attached to T; Double line between N and C

Represents a single bond or a double bond, provided that when it is a double bond, R ₉ is absent and R ₁₀ is H, and when it is a single bond, R ₉ is H or is selected from acetyl, trifluoroacetyl, uncle An amino-protecting moiety of butoxycarbonyl, benzyloxycarbonyl and 9-fluorenylmethoxycarbonyl; R _{10 is} selected from OH, an ether represented by O R ₁₁ , sulfite SO ^{3 -} or OSO ^3- , wherein R _{11 is} selected from the group consisting of 1 a linear, branched or cyclic alkyl, alkenyl or alkynyl group of -10 carbon atoms; The dashed line between C2 and C3 indicates the optional presence of a double bond; When there is a double bond between C2 and C3, R _{12 is} selected from C _5-10 aryl optionally substituted by one or more substituents, wherein the substituent is selected from the group consisting of halogen, nitro, cyano, amino, ether, carboxyl, ester, C _{a 1-7} hydrocarbon group, a C _3-7 heterocyclic group; When there is a single bond between C2 and C3, R ₁₂ is

Wherein R ₁₄ and R _{15 are} independently selected from the group consisting of H, F, C _1-4 saturated alkyl, C _2-3 alkenyl, the alkyl and alkenyl optionally substituted by a group selected from the group consisting of a C _1-4 alkyl amide group and a C _1-4 alkyl ester, or, when one of R ₁₄ and R ₁₅ is H, the other is selected from the group consisting of a nitrile and a C _1-4 alkyl ester; Module D1" may be substituted by one or more R ₁₃ wherein R _{13 is} selected from the group consisting of H, halogen, nitro, cyano, amino, ether, carboxyl, ester, C _1-7 hydrocarbyl, C _3-7 heterocyclyl . The cytotoxic drug according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein R _{3 is} selected from the group consisting of H, OH, SH, NH ₂ , N ₃ , NO ₂ , NO , CF ₃ , CN, C(O)NH ₂ , C(O)H, C(O)OH, halogen, ^Rh , SR ^h , S(O)R ^h , S(O) ₂ R ^h , S( O) OR ^h , S(O) ₂ OR ^h , OS(O)R ^h , OS(O) ₂ R ^h , OS(O)OR ^h , OS(O) ₂ OR ^h , OR ^h , NHR ^h , N (R ^h )R ⁱ , ⁺ N(R ^h )(R ⁱ )R ^j , P(O)(OR ^h )(OR ⁱ ), OP(O)(OR ^h )(OR ⁱ ), SiR ^h R ⁱ R ^j , C(O)R ^h , C(O)OR ^h , C(O)N(R ^h )R ⁱ , OC(O)R ^h , OC(O)OR ^h , OC(O)N(R ^h ) R ⁱ , N(R ^h )C(O)R ⁱ , N(R ^h )C(O)OR ⁱ or N(R ^h )C(O)N(R ⁱ )R ^j ; Wherein R ^h , R ⁱ and R ^{j are} independently selected from H and optionally substituted C _1-15 alkyl, C _1-15 heteroalkyl, C _3-15 cycloalkyl, C _1-15 heterocycloalkane a C _5-15 aryl group or a C _1-15 heteroaryl group; T is selected from a C ₂ -C ₁₂ hydrocarbyl group, Z, (C ₁ -C ₆ alkylene)-Z-(C ₁ -C ₆ alkylene), (C ₁ -C ₆ alkylene)-Z- (C ₁ -C ₆ alkylene)-Z-(C ₁ -C ₆ alkylene), (C ₁ -C ₆ alkenylene)-Z-(C ₁ -C ₆ alkenylene) or (C _1- C ₆ alkynylene)-Z-(C ₁ -C ₆ alkynylene); Wherein Z is independently selected from the group consisting of O, S, NR ₁ , aryl and heteroaryl; Wherein the alkylene, alkenylene, aryl and heteroaryl groups are independent and optionally F, OH, O(C ₁ -C ₆ alkyl), NH ₂ , NHCH ₃ , N(CH ₃ ) ₂ and C ₁ -C ₆ alkyl substituted wherein the alkyl group is optionally substituted with one or more F. ; The cytotoxic drug according to claim 1 or 2, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein X ₁ is O and R ₁ is H; The cytotoxic drug or a pharmaceutically acceptable salt, hydrate or solvate thereof according to claim 1 or 2, wherein D1 is D1', a double bond exists between N and C, and C2 and C3 are present. a double bond is present, and R ₁₂ is a methoxy substituted phenyl group; Pharmaceutically claim 12 or cytotoxic agents or a pharmaceutically acceptable salt, hydrate or solvate thereof as claimed in claim, characterized in that, where D1 is D2 ", the presence of a double bond, and R is between C ₁₃ and N Methoxy The cytotoxic drug or a pharmaceutically acceptable salt, hydrate or solvate thereof according to claim 1 or 2, wherein T is selected from the group consisting of C2-12 alkylene groups. The cytotoxic drug or a pharmaceutically acceptable salt, hydrate or solvate thereof according to claim 1 or 2, wherein T is

A pharmaceutical composition comprising the cytotoxic drug according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier, a glidant, a diluent , or an excipient. A cytotoxic drug according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment of a tumor, an autoimmune disease or an infectious disease.

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