CN106188098A - A kind of hydridization cancer therapy drug and preparation method and application - Google Patents

Abstract

The invention discloses a hybrid anticancer drug and its preparation method and application, belonging to the technical field of medicine. The structural formula of the hybrid anticancer drug of the present invention is shown in formula I: wherein: X is selected from one of -SCH ₂ -, -CH=CH- or -(CH ₂ ) ₂ -substituents; Y is selected from One of -O- or -OCH ₂ O-; R ₂ is selected from one of -CN or -ONO ₂ . The hybrid anticancer drug designed and synthesized by the present invention has strong binding affinity with XIAP-BIR3, promotes cancer cell apoptosis, has more obvious non-peptide features, can act on multiple targets, and has multiple effects.

Description

A kind of hybrid anticancer drug and its preparation method and application

technical field

The invention belongs to the technical field of medicine, in particular to a hybrid anticancer drug and its preparation method and application.

Background technique

In the past few years, in order to make drugs have dual or multiple effects, hybrid drugs that hybridize two pharmacophores into one drug molecule have become a new hot spot in the development of new drugs. Due to the complexity of many serious diseases such as cancer and neurodegenerative diseases, these diseases are difficult to be cured by a highly selective pharmacophore, which is why hybrid drugs have received special attention in recent years. Compared with ordinary drugs, hybrid drugs can be used to improve molecular affinity and selectivity, reduce side effects, etc. The constituent units of hybrid molecules can be artificially designed or natural organic small molecules, polypeptides, nucleic acids, etc. For the target, these pharmacophores are expected to play a synergistic role.

Apoptosis is a programmed cell death behavior regulated by a series of orderly signal flows in multicellular organisms. It is a normal physiological process for the body to eliminate harmful cells and prevent excessive cell proliferation. plays an important role in. Studies have shown that dysregulation of apoptosis mechanism is an important pathological feature of diseases such as cancer. In the past few decades, although some progress has been made in cancer treatment, the tolerance of tumor cells to various cytotoxic stimuli and the disorder and dysregulation of apoptosis are still the main obstacles that cancer treatment has to overcome.

Inhibitor of apoptosis (IAP) is a key regulator of cell death in the apoptosis pathway. Inhibitors of apoptosis are overexpressed in a variety of tumor cells. At present, the IAPs family includes eight members, namely Cp-IAP, Op-IAP, XIAP, c-IAP1, c-IAP2, NAIP, Livin/ML-IAP and Survivin, among which four apoptosis inhibitory proteins XIAP, c- IAP1, c-IAP2 and ML-IAP are directly involved in the regulation of apoptosis, and these IAPs proteins inhibit apoptosis mainly by inhibiting caspase.

Smac/DIABLO (a second mitochondrial-derived cystinease activator/low isoelectric point IAP direct binding protein) is an endogenous antagonist of IAP released from mitochondria to the cytoplasm. Smac binds to IAPs through its N-terminal Ala-Val-Pro-Ile (AVPI) tetrapeptide motif, thereby removing the inhibitory effect of IAPs on cell apoptosis and promoting cell apoptosis. Using AVPI tetrapeptide molecule as the lead compound, designing and synthesizing non-peptide small molecule Smac mimics, targeting IAPs, and developing anticancer drugs that induce apoptosis have become a research hotspot in the field of cancer treatment.

Contents of the invention

In order to overcome the shortcomings and deficiencies in the above-mentioned prior art, the primary purpose of the present invention is to provide a hybrid anticancer drug. The hybrid anticancer drug can antagonize the effect of cell apoptosis inhibitory protein and produce quinone methide intermediate.

Another object of the present invention is to provide a preparation method of the above-mentioned hybrid anticancer drug.

Another object of the present invention is to provide the application of the above-mentioned hybrid anticancer drug.

The object of the present invention is achieved through the following technical scheme: a hybrid anticancer drug, its structural formula is as shown in formula I:

in:

X is selected from one of -SCH ₂ -, -CH═CH- or -(CH ₂ ) ₂ - substituents:

Y is selected from one of -O- or -OCH ₂ O-;

_{R1 is} selected from one of the substituents;

R ₂ is selected from one of -CN or -ONO ₂ .

The preparation method of the above-mentioned hybrid anticancer drug comprises the following steps:

1) Dissolve compound A and inorganic base in a polar organic solvent, stir the suspension for 1 to 30 minutes, then add methyl iodide or ethyl iodide, stir the mixture at room temperature for 4 to 12 hours under the protection of an inert gas, and concentrate , the resulting residue was dissolved in a non-polar organic solvent, the organic phase was washed with water, dried with a desiccant, and the crude product obtained by concentration was purified by column to obtain a colorless oily substance B;

Wherein, the molar ratio of compound A, inorganic base, methyl iodide or ethyl iodide is: 1:1-10:1-5;

2) At -10-15°C, add the chlorinated hydrocarbon solution of TFA to the chlorinated hydrocarbon solution of compound B, stir the mixture for 0.3-5 hours, and concentrate to obtain the crude product amine C. At -10-15°C, add Dissolve compound C in a dry chlorinated hydrocarbon solution, add tert-butoxycarbonyl-protected amino acid, condensing agent and additives, then add an organic base to adjust the pH of the reaction solution to alkaline, stir the mixture at room temperature overnight, and then wash the reaction solution with dilute acid solution, dilute alkali solution, saturated brine, dried, and concentrated under reduced pressure to obtain a crude product that was purified by column to obtain a colorless oily substance D;

Wherein, the molar ratio of compound C, tert-butoxycarbonyl-protected amino acid, condensing agent and additive is 1:1-5:1-20:0.25-20;

3) Dissolve compound D in a mixed solvent of polar organic solvent and water, add a strong base, stir the suspension at room temperature for 2 to 7 hours, concentrate, dilute the residue with water, adjust the pH value to acidic with dilute hydrochloric acid solution, and use The non-polar organic solvent was extracted three times, the organic phases were combined, dried with a desiccant, and concentrated to obtain a colorless oil E;

Wherein, the molar ratio of the compound D to the strong base is 1:1-5;

4) At -10-15°C, dissolve compound E in a dry chlorinated hydrocarbon solution, add an organic base or a mixed chlorinated hydrocarbon solution of a condensing agent and an organic base, and then add 2-(4-(chloromethoxy )phenyl)acetonitrile or 2-(4-(chloromethoxy)phenyl)methyl nitrate or 2-(4-hydroxyphenyl)acetonitrile or 2-(4-hydroxyphenyl)methyl nitrate , the mixed solution was stirred overnight at room temperature under the protection of an inert gas, and concentrated to obtain the crude product F. At -10 to 15°C, TFA was added to the chlorinated hydrocarbon solution of compound F, the mixed solution was stirred for 0.1 to 5 hours, concentrated, and the high-efficiency liquid Phase purification, freeze-drying to obtain white finished product G;

Wherein, the compound E, 2-(4-(chloromethoxy)phenyl)acetonitrile or 2-(4-(chloromethoxy)phenyl)methyl nitrate or 2-(4- The molar ratio of hydroxyphenyl) acetonitrile or 2-(4-hydroxyphenyl)methyl nitrate, organic base and condensing agent is 1:1～5:1～20:1～20.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the general structural formula of the compound A is: Wherein, X is one of -SCH ₂ -, -CH═CH- or -(CH ₂ ) ₂ -.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the inorganic base is one or more of K ₂ CO ₃ , Na ₂ CO ₃ , KHCO ₃ , NaHCO ₃ , KOH, NaOH or LiOH.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the strong base is one or more of KOH, NaOH, Ca(OH) ₂ , NaH, KH or LiOH.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the polar organic solvent is one or more of DMF, DMSO, THF or MeCN.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the non-polar organic solvent is one or more of ethyl acetate, dichloromethane, chloroform or ether.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the inert gas is nitrogen or argon.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the chlorinated hydrocarbon solution is dichloromethane or chloroform.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the amino acid protected by the tert-butoxycarbonyl group is N-tert-butoxycarbonyl-L-alanine, (S)-2-(tert-butoxycarbonyl) One or more of aminobutyric acid, (S)-2-((tert-butoxycarbonyl)methylamino)propionic acid or (S)-2-((tert-butoxycarbonyl)methylamino)butanoic acid .

Preferably, in the preparation method of the above hybrid anticancer drug, the condensing agent is one or more of BOP, DCC, EDCI, DIC, PyBOP, TBTU, HBTU, HATU or TATU.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the additive is one or more of HOBt, HOOBt or HOSu.

Preferably, in the preparation method of the above-mentioned hybrid anticancer drug, the organic base is one or more of triethylamine, pyridine, DBU, DIEA or N-methylmorpholine.

Preferably, in the preparation method of the above hybrid anticancer drug, the desiccant is one of anhydrous magnesium sulfate, anhydrous sodium sulfate, anhydrous potassium carbonate or anhydrous calcium chloride.

The application of the above-mentioned hybrid anticancer drug as an anticancer drug. Compared with the prior art, the technical solution provided by the present invention has stronger binding affinity with XIAP-BIR3, more obvious non-peptide features, and is expected to act on multiple anti-cancer targets.

Compared with the prior art, the present invention has the following advantages and effects:

In order to strengthen the anticancer activity of Smac mimics, the present invention replaces the hydrophobic group at the carbon end of Smac mimics with a new group, and designs and synthesizes a class of hybrid anticancer drugs. The introduced new group not only interacts with the apoptosis inhibitory protein, but also generates quinone methide under the hydrolysis of esterase. The methide intermediates of quinones are highly electrophilic and can participate in many physiological processes, inducing cell death and inhibiting cell growth. Therefore, this type of hybrid drug includes two pharmacophores: one pharmacophore binds to IAPs and antagonizes the inhibitory effect of IAPs on cell apoptosis; the other pharmacophore produces quinone methide intermediates and participates in multiple physiological process. Therefore, the hybrid anticancer drug designed and synthesized by the present invention has stronger binding affinity with XIAP-BIR3, more obvious non-peptide features, can act on multiple targets, and play multiple functions.

Description of drawings

Fig. ₁ is the ¹ H NMR detection spectrogram of compound G1;

Fig. ₂ is the ¹ H NMR detection spectrogram of compound G2;

Fig. ₃ is the ¹ H NMR detection spectrogram of compound G3;

Figure 4 is a competitive binding inhibition curve between compound G ₁ and XIAP-BIR3;

Fig. 5 shows the synergistic effect of compound G ₁ and TRAIL on the apoptosis of MDA-MB-231 cells.

detailed description

The claims of the present invention will be further described in detail below in conjunction with specific embodiments, but this does not constitute any restriction on the present invention. Anyone who makes limited modifications within the scope of the claims of the present invention is still within the rights of the present invention. within the scope of protection.

All reagents were commercially available analytically pure unless otherwise specified, and were used directly without further purification. The glass instruments for the anhydrous and oxygen-free reaction were dried in an oven before use, and the reaction was carried out under the protection of Ar. Solvent redistillation was carried out under the protection of Ar, THF was dried under the condition of metallic sodium and benzophenone, and dichloromethane and ethyl acetate were dried with calcium hydride. Use 63-200 mesh silica gel for column chromatography, and Merck TLC silica gel 60F254 plastic chromatographic plate for thin layer chromatography, and use 254nm ultraviolet light, iodine vapor, or 2% ethanol solution of ninhydrin for color detection. The nuclear magnetic resonance spectrum was measured using a Bruker AVANCE III400NMR spectrometer, and the chemical shift values were recorded in ppm. Mass spectra were determined using an LCT Premier XE time-of-flight mass spectrometer. Optical rotation values were measured with a JASCO P-1010 polarimeter.

High performance liquid chromatography (HPLC) conditions: (1) Preparative HPLC: Perkin Elmer Series 200UV/VIS detector; Perkin Elmer Series 200 two-phase pump; vacuum degasser; Gilson FC203b fraction receiver; (2) Analytical HPLC : Perkin Elmer Series 225 autosampler; Perkin Elmer Series 220 UV/VIS detector; Perkin Elmer Series 200 four-phase pump; vacuum degasser. Mobile phase: (1) Mobile phase A: water+0.045% TFA; (2) Mobile phase B: 10% water+90% acetonitrile+0.038% TFA.

Example 1 (hybrid anticancer drug G ₁ )

The synthesis method of compound A ₁ (the structural formula of A ₁ is shown in formula II) refers to the preparation method in paragraphs [0046] to [0083] of Chinese patent 201610036840.2.

(5R,8S,10aR)-5-((tert-butoxycarboxamido)-6-carbonylthiazepine[1,5]pyrrolidine[2,1-d]-8-carboxylic acid methyl ester Preparation of (Compound B ₁ ):

Compound A ₁ (36 mg, 0.10 mmol) and K ₂ CO ₃ (69 mg, 0.50 mmol) were dissolved in DMF (2 mL). The suspension was stirred for 15 minutes, then iodomethane (31 μL, 0.50 mmol) was added. The mixture was stirred at room temperature for 8 hours under the protection of Ar, concentrated, and the resulting residue was dissolved in ethyl acetate. The organic phase was washed with water, dried over anhydrous magnesium sulfate, concentrated and the resulting crude product was purified by column (cyclohexane/ethyl acetate=3/1) to obtain a colorless oily substance B ₁ (28mg, 78%), the reaction chemical equation is shown in Formula II shown.

¹ H NMR (400MHz, CDCl ₃ ): δ5.87(d, J=6.9Hz, 1H), 4.71-4.66(m, 1H), 4.61-4.56(m, 1H), 4.49(t, J=9.0Hz ,1H),3.74(s,3H),2.96-2.88(m,2H),2.75-2.67(m,2H),2.43-2.36(m,1H),2.14-1.91(m,3H),1.80(dd , J＝11.7, 6.7Hz, 1H), 1.75-1.66(m, 1H), 1.40(s, 9H); ¹³ C NMR (100MHz, CDCl ₃ ): δ173.29, 170.41, 154.97, 80.06, 59.67, 56.82, 56.34 ,52.63,38.43,31.40,30.49,28.64,27.04.

(5R,8S,10aR)-5-((S)-2-(tert-butoxycarboxamido)propionamido)-6-carbonylthiazepine[1,5]pyrrolidine[2,1 -d] - Preparation of methyl 8-carboxylate (compound D ₁ ):

A solution of TFA (300 μL) in dichloromethane (1 mL) was added to a solution of compound B ₁ (28 mg, 0.08 mmol) in dichloromethane (2 mL) at -10°C. The mixture was stirred for 5 h and concentrated to obtain compound amine C ₁ . At -10°C, the obtained compound C ₁ was dissolved in dry dichloromethane (3 mL), and Boc-Ala-OH (15 mg, 0.08 mmol), EDCI (307 mg, 1.6 mmol), HOBt (216 mg, 1.6 mmol) were added ), then add DIEA to adjust the reaction solution to alkaline. The mixture was stirred overnight at room temperature, and the reaction solution was washed successively with dilute hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine. The organic phase was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and the resulting crude product was purified by column (cyclohexane/acetone=3/1) to obtain a colorless oil D ₁ (25 mg, the total yield of the two-step reaction was 73%) , the reaction chemical equation is shown in formula Ⅲ.

¹ H NMR (400MHz, CDCl ₃ ): δ7.33(d, J=4.2Hz, 1H), 5.02(d, J=6.1Hz, 1H), 4.81-4.76(m, 1H), 4.72-4.67(m ,1H),4.49(t,J=9.0Hz,1H),4.19-4.05(m,1H),3.74(s,3H),2.98-2.86(m,2H),2.76-2.65(m,2H), 2.44-2.37(m,1H),2.15-1.92(m,3H),1.81(dd,J=11.8,6.8Hz,1H),1.72(tt,J=13.9,4.5Hz,1H),1.42(s, 9H), 1.32 (d, J=7.1Hz, 3H); ¹³ CNMR (100MHz, CDCl ₃ ): δ173.18, 171.85, 169.90, 155.52, 80.38, 59.72, 56.39, 55.76, 52.67, 50.63, 38.01, 37.09, 31.54, 31.38, 28.62, 27.02, 19.13.

(5R,8S,10aR)-5-((S)-2-(tert-butoxycarboxamido)propionamido)-6-carbonylthiazepine[1,5]pyrrolidine[2,1 Preparation of -d]-8-carboxylic acid (compound E ₁ ):

Compound D ₁ (25 mg, 0.06 mmol) was dissolved in a mixed solvent of MeCN/H ₂ O (1:1, 2 mL), and KOH (3.3 mg, 0.06 mmol) was added. The suspension was stirred at room temperature for 2 hours, concentrated, the residue was diluted with water, and the pH was adjusted to 2 with dilute hydrochloric acid solution. The aqueous solution was extracted three times with ethyl acetate, the organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to obtain a colorless oil E ₁ (23 mg, 95%). The next reaction was carried out directly without further purification. The reaction chemical equation is shown in Formula IV.

¹ H NMR (400MHz, MeOD): δ4.84(dd, J=10.1, 2.3Hz, 1H), 4.76-4.71(m, 1H), 4.48(t, J=8.8Hz, 1H), 4.08(q, J＝7.2Hz, 1H), 2.98-2.72(m, 4H), 2.49-2.43(m, 1H), 2.20-2.00(m, 3H), 1.86(dd, J＝11.1, 6.1Hz, 1H), 1.80 -1.72(m,1H),1.46(s,9H),1.33(d,J=7.2Hz,3H); ¹³ C NMR(100MHz,MeOD):δ177.10,175.63,172.01,158.56,81.59,62.07,58.77, 57.62, 52.57, 38.94, 38.71, 32.69, 32.16, 29.55, 28.76, 18.87.

(5R,8S,10aR)-(4-(cyanomethyl)phenoxy)methyl 5-((S)-2-aminopropionamido)-6-carbonylthiazepine[1,5] Preparation of pyrrolidine[2,1-d]-8-carboxylate trifluoroacetate (G ₁ ):

At -10°C, compound E ₁ (23mg, 0.06mmol) was dissolved in dry dichloromethane solution (3mL), triethylamine (8μL, 0.06mmol) was added, and then 2-(4-(chloromethoxy (1) phenyl) acetonitrile (11 mg, 0.06 mmol) in dry dichloromethane (2 mL). The mixture was stirred overnight at room temperature under the protection of Ar, and concentrated to obtain the crude product F ₁ . Compound F1 was dissolved in dichloromethane ( ₂ mL) at -10°C, TFA (300 μL) in dichloromethane solution (1 mL) was added, the mixture was stirred for 5 hours, the resulting residue was concentrated, purified by HPLC, and lyophilized. A white solid G ₁ was obtained (7.8 mg, 23% over two steps). The reaction chemical equation is shown in Formula V. The ¹ H NMR detection spectrum of compound G ₁ is shown in FIG. 1 .

¹ H NMR (400MHz, D ₂ O): δ7.27 (d, J = 8.8Hz, 2H), 7.01 (d, J = 8.8Hz, 2H), 5.84, 5.76 (ABq, J = 6.8Hz, 2H) ,4.49-4.44(m,1H),4.35(t,J=9.1Hz,1H),3.97(q,J=7.1Hz,1H),3.76(s,2H),2.76(dd,J=14.1,3.0 Hz,1H),2.67(dt,J＝15.6,4.5Hz,1H),2.32-2.24(m,3H),2.07-1.97(m,1H),1.93-1.83(m,1H),1.72-1.62( m,3H),1.36(d,J＝7.1Hz,3H); ¹³ CNMR(100MHz,D ₂ O):δ172.82,172.77,170.92,170.88,169.81,169.77,155.11,155.05,129.72,129.67,125.71,125.63 ,120.19,120.13,117.22,117.14,86.23,86.15,60.62,60.57,57.94,57.88,54.70,54.65,48.91,48.83,35.94,35.88,34.73,34.66,30.89,30.83,29.48,29.42,26.47,26.41,22.03 , 21.98, 16.44, 16.39; MS (ESI): m/z 461.2 (M+H) ⁺ ; HR ESI MS for C ₂₂ H ₂₉ N ₄ O ₅ S required 461.1859, found: 461.1880.

Example 2 (hybrid anticancer drug G ₂ )

The synthesis method of compound A ₂ (the structural formula of A ₂ is shown in formula VI) refers to the method reported by Duggan, HM et al. in Organic & biomolecular chemistry ("Organic and Biomolecular Chemistry"), Issue 3, 2005, page 2287.

(3S,6S,10aR)-6-(tert-butoxycarboxamido)-5-carbonyl-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a] Preparation of ethyl azacyclooctene-3-carboxylate (compound B ₂ ):

Compound A ₂ (32 mg, 0.10 mmol) and Na ₂ CO ₃ (106 mg, 1.00 mmol) were dissolved in DMSO (2 mL). The suspension was stirred for 30 minutes, then ethyl iodide (8 μL, 0.10 mmol) was added. The mixture was stirred at room temperature for 4 hours under N ₂ protection, concentrated, and the resulting residue was dissolved in dichloromethane. The organic phase was washed with water, dried over anhydrous sodium sulfate, and the crude product was concentrated and purified by column (cyclohexane/ethyl acetate=6/1) to obtain a colorless oil B ₂ (25 mg, 72%). The reaction chemical equation is shown in Formula VI.

¹ H NMR (400MHz, CDCl ₃ ): δ5.83-5.72(m, 1H), 5.72-5.62(m, 1H), 5.56(d, J=7.5Hz, 1H), 4.86-4.76(m, 1H) ,4.50-4.39(m,1H),4.12(q,J＝7.1Hz,3H,containing 1H),2.76-2.70(m,2H),2.45-2.33(m,1H),2.32-2.19(m,1H ), 2.15-2.06(m, 1H), 2.06-1.98(m, 1H), 1.98-1.82(m, 2H), 1.39(s, 9H), 1.22(t, J=7.1Hz, 3H); 13C NMR (100MHz, CDCl3): δ171.89, 171.05, 155.23, 129.32, 125.92, 79.58, 60.97, 60.38, 58.74, 51.91, 35.37, 33.03, 32.97, 28.42 (3C), 27.29, 14.21.

(3S,6S,10aR)-6-((S)-2-(tert-butoxycarboxamido)propionylamino)-5-carbonyl-1,2,3,5,6,7,10,10a- Preparation of ethyl octahydropyrrolo[1,2-a]azacyclooctene-3-carboxylate (compound D ₂ ):

A solution of TFA (300 μL) in dichloromethane (1 mL) was added to a solution of compound B ₂ (28 mg, 0.08 mmol) in dichloromethane (2 mL) at 0 °C. The mixture was stirred for 2.5 h and concentrated to obtain compound amine C ₂ . At 0°C, the obtained compound C2 was dissolved in dry dichloromethane ( ₃ mL), and Boc-Ala-OH (38 mg, 0.20 mmol), BOP (35 mg, 0.08 mmol), HOOBt (3 mg, 0.02 mmol) were added , and then add NEt ₃ to adjust the reaction solution to alkaline. The mixture was stirred overnight at room temperature, and the reaction solution was washed successively with saturated ammonium chloride solution, saturated potassium bicarbonate solution and saturated brine. The organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained crude product was purified by column (cyclohexane/ethyl acetate=5/1) to obtain colorless oily substance D ₂ (26 mg, total yield of two-step reaction 77 %). The reaction chemical equation is shown in Formula VII.

¹ H NMR (400MHz, CDCl ₃ ): δ7.06(d, J=5.7Hz, 1H), 5.93-5.78(m, 1H), 5.78-5.62(m, 1H), 5.13-4.90(m, 2H) ,4.46(dd,J＝8.9,3.9Hz,1H),4.28-4.05(m,4H),2.87-2.66(m,2H),2.54-2.38(m,1H),2.38-2.24(m,1H) ,2.24-2.08(m,1H),2.08-1.82(m,3H),1.43(s,9H),1.33(d,J=7.0Hz,3H),1.26(t,J=7.1Hz,3H).

(3S,6S,10aR)-6-((S)-2-(tert-butoxycarboxamido)propionylamino)-5-carbonyl-1,2,3,5,6,7,10,10a- Preparation of Octahydropyrrolo[1,2-a]azacyclooctene-3-carboxylic acid (Compound E ₂ ):

Compound D ₂ (25 mg, 0.06 mmol) was dissolved in a mixed solvent of THF/H ₂ O (1:1, 2 mL), and LiOH (4.3 mg, 0.18 mmol) was added. The suspension was stirred at room temperature for 4.5 hours, concentrated, the residue was diluted with water, and the pH was adjusted to 2 with dilute hydrochloric acid solution. The aqueous solution was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain a colorless oil E ₂ (22 mg, 93%). The next reaction was carried out directly without further purification. The reaction chemical equation is shown in Formula VIII.

¹ H NMR (400MHz, CDCl ₃ ): δ9.55(bs,1H),7.63(bs,1H),7.50(bs,1H),5.96-5.62(m,2H),5.35-5.14(m,1H) ,4.71-4.45(m,1H),4.33-4.08(m,2H),3.00-2.70(m,2H),2.54-2.32(m,1H),2.32-2.14(m,2H),2.14-1.98( m, 2H), 1.98-1.84 (m, 1H), 1.44 (d, J=5.0Hz, 9H), 1.37 (dd, J=6.9, 3.5Hz, 3H); ¹³ C NMR (100MHz, CDCl ₃ ): δ172.33, 155.85, 129.25, 125.94, 80.54, 60.73, 59.29, 50.52, 49.93, 34.59, 33.07, 32.78, 28.46, 26.59, 19.56, 14.33.

(3S,6S,10aR)-(4-ethylcyanophenyl)-6-((S)-2-aminopropionamido)-5-carbonyl-1,2,3,5,6,7,10 , Preparation of 10a-octahydropyrrolo[1,2-a]azacycloctane-3-carboxylate trifluoroacetate (G ₂ ):

At 0°C, compound E ₂ (22 mg, 0.06 mmol) was dissolved in dry chloroform (5 mL), a mixed chloroform solution of EDCI (115 mg, 0.60 mmol) and DIEA (99 μL, 0.60 mmol) was added, and then 2-( 4-Hydroxyphenyl) acetonitrile (24 mg, 0.18 mmol), and the mixture was stirred at room temperature overnight. The solvent was dried under reduced pressure to obtain F ₂ as a pale yellow oil. F2 was dissolved in chloroform (3mL), cooled to ₀ °C, TFA (0.5mL) was added dropwise, after 2.5h, the solvent was evaporated to dryness, the residue was concentrated, purified by HPLC, and freeze-dried to obtain a white solid G2 ₍ 3.1mg , the total yield of the two-step reaction was 10%). The reaction chemical equation is shown in Formula IX. The ¹ HNMR detection spectrum of compound G ₂ is shown in FIG. 2 .

¹ H NMR (400MHz, CDCl ₃ ): δ7.70(bs, 1H), 7.32(d, J=8.3Hz, 2H), 7.07(d, J=8.3Hz, 2H), 5.85-5.51(m, 2H ),5.16-5.04(m,1H),4.64(d,J＝5.9Hz,1H),4.33-4.16(m,1H),4.16-3.99(m,1H),3.72(s,2H),2.86- 2.59(d, J=14.8Hz, 2H), 2.51-2.34(m, 1H), 2.25-2.20(m, 2H), 2.20-2.07(m, 3H), 2.06-1.90(m, 2H), 1.55- 1.43(m,3H). MS(ESI): m/z 411.2(M+H) ⁺ ; HR ESI MS for C ₂₂ H ₂₇ N ₄ O ₄ required 411.1954, found: 411.1932.

Example 3 (hybrid anticancer drug G ₃ )

The synthesis method of compound A ₂ (the structural formula of A ₂ is shown in formula X) refers to the method reported by Duggan, HM et al. in Organic & biomolecular chemistry ("Organic and Biomolecular Chemistry"), No. 3, 2005, page 2287.

(3S,6S,10aR)-6-(tert-butoxycarboxamido)-5-carbonyl-1,2,3,5,6,7,10,10a-octahydropyrrolo[1,2-a] Preparation of ethyl azacyclooctene-3-carboxylate (compound B ₂ ):

Compound A ₂ (32 mg, 0.10 mmol) and KOH (6 mg, 0.10 mmol) were dissolved in THF (2 mL). The suspension was stirred for 1 min, then ethyl iodide (20 μL, 0.25 mmol) was added. The mixture was stirred at room temperature for 12 hours under N ₂ protection, concentrated, and the obtained residue was dissolved in chloroform. The organic phase was washed with water, dried over anhydrous potassium carbonate, and the crude product was concentrated and purified by column (cyclohexane/ethyl acetate=6/1) to obtain a colorless oil B ₂ (29 mg, 82%). The reaction chemical equation is shown in Formula X.

¹ H NMR (400MHz, CDCl ₃ ): δ5.83-5.72(m, 1H), 5.72-5.62(m, 1H), 5.56(d, J=7.5Hz, 1H), 4.86-4.76(m, 1H) ,4.50-4.39(m,1H),4.12(q,J＝7.1Hz,3H,containing 1H),2.76-2.70(m,2H),2.45-2.33(m,1H),2.32-2.19(m,1H ), 2.15-2.06(m, 1H), 2.06-1.98(m, 1H), 1.98-1.82(m, 2H), 1.39(s, 9H), 1.22(t, J=7.1Hz, 3H); 13C NMR (100MHz, CDCl3): δ171.89, 171.05, 155.23, 129.32, 125.92, 79.58, 60.97, 60.38, 58.74, 51.91, 35.37, 33.03, 32.97, 28.42 (3C), 27.29, 14.21.

(3S,6S,10aR)-6-((S)-2-(tert-butoxycarboxamido)propionylamino)-5-carbonyl-1,2,3,5,6,7,10,10a- Preparation of ethyl octahydropyrrolo[1,2-a]azacyclooctene-3-carboxylate (compound D ₂ ):

A chloroform solution (1 mL) of TFA (300 μL) was added to a chloroform solution (2 mL) of compound B ₂ (28 mg, 0.08 mmol) at 15°C. The mixture was stirred for 0.3 h and concentrated to obtain compound amine C ₂ . At 15°C, the obtained compound C2 was dissolved in dry chloroform ( ₃ mL), and Boc-Ala-OH (76 mg, 0.40 mmol), DCC (165 mg, 0.80 mmol), HOSu (92 mg, 0.80 mmol) were added, and then Add DBU to adjust the pH of the reaction solution to alkaline. The mixture was stirred overnight at room temperature, and the reaction solution was washed successively with saturated ammonium chloride solution, saturated potassium bicarbonate solution and saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was purified by column (cyclohexane/ethyl acetate=5/1) to obtain a colorless oily substance D ₂ (23 mg, the total yield of the two-step reaction was 68 %). The reaction chemical equation is shown in formula Ⅺ.

¹ H NMR (400MHz, CDCl ₃ ): δ7.06(d, J=5.7Hz, 1H), 5.93-5.78(m, 1H), 5.78-5.62(m, 1H), 5.13-4.90(m, 2H) ,4.46(dd,J＝8.9,3.9Hz,1H),4.28-4.05(m,4H),2.87-2.66(m,2H),2.54-2.38(m,1H),2.38-2.24(m,1H) ,2.24-2.08(m,1H),2.08-1.82(m,3H),1.43(s,9H),1.33(d,J=7.0Hz,3H),1.26(t,J=7.1Hz,3H).

(3S,6S,10aR)-6-((S)-2-(tert-butoxycarboxamido)propionylamino)-5-carbonyl-1,2,3,5,6,7,10,10a- Preparation of Octahydropyrrolo[1,2-a]azacyclooctene-3-carboxylic acid (Compound E ₂ ):

Compound D ₂ (25 mg, 0.06 mmol) was dissolved in a mixed solvent of MeCN/H ₂ O (1:1, 2 mL), and NaOH (12 mg, 0.30 mmol) was added. The suspension was stirred at room temperature for 7 hours, concentrated, the residue was diluted with water, and the pH was adjusted to 2 with dilute hydrochloric acid solution. The aqueous solution was extracted three times with chloroform, the organic phases were combined, dried over anhydrous potassium carbonate, and concentrated to obtain a colorless oil E ₂ (20 mg, 84%). The next reaction was carried out directly without further purification. The reaction chemical equation is shown in Formula XII.

¹ H NMR (400MHz, CDCl ₃ ): δ9.55(bs,1H),7.63(bs,1H),7.50(bs,1H),5.96-5.62(m,2H),5.35-5.14(m,1H) ,4.71-4.45(m,1H),4.33-4.08(m,2H),3.00-2.70(m,2H),2.54-2.32(m,1H),2.32-2.14(m,2H),2.14-1.98( m, 2H), 1.98-1.84 (m, 1H), 1.44 (d, J=5.0Hz, 9H), 1.37 (dd, J=6.9, 3.5Hz, 3H); ¹³ C NMR (100MHz, CDCl ₃ ): δ172.33, 155.85, 129.25, 125.94, 80.54, 60.73, 59.29, 50.52, 49.93, 34.59, 33.07, 32.78, 28.46, 26.59, 19.56, 14.33.

(3S,6S,10aR)-(4-Nitrate methylenephenyl)-6-((S)-2-Aminopropionamido)-5-carbonyl-1,2,3,5,6,7 , Preparation of 10,10a-octahydropyrrolo[1,2-a]azacycloctane-3-carboxylate trifluoroacetate (G ₃ ):

At 15°C, compound E ₂ (20 mg, 0.05 mmol) was dissolved in dry chloroform (5 mL), a mixed chloroform solution of DCC (206 mg, 1 mmol) and DIEA (165 μL, 1 mmol) was added, and then 2-(4-hydroxy Phenyl)methyl nitrate (42mg, 0.25mmol), the mixture was stirred at room temperature overnight. The solvent was dried under reduced pressure to obtain F ₃ as a pale yellow oil. F ₃ was dissolved in chloroform (3 mL), and TFA (0.5 mL) was added dropwise at 15°C. After 0.3 h, the solvent was evaporated to dryness, and the resulting residue was purified by HPLC and freeze-dried to obtain a white solid G ₃ (2.3 mg ,8%). The reaction chemical equation is shown in Formula XIII. The ¹ H NMR detection spectrum of compound G ₃ is shown in FIG. 3 .

¹ H NMR (400MHz, CDCl ₃ ): δ7.46-7.29(m,2H),7.27-7.10(m,2H),5.95-5.66(m,2H),5.04-4.80(m,1H),4.67( dd,J＝7.7,6.1Hz,1H),4.24-4.10(m,2H),4.07-3.99(m,1H),3.71-3.51(m,1H),2.77-2.47(m,2H),2.47- 2.23(m,1H),2.20-1.45(m,7H),1.54(d,J=6.4Hz,3H).MS(ESI):m/z 447.2(M+H) ⁺ ; HR ESI MS for C ₂₁ H ₂₇ N ₄ O ₇ required 446.1801, found: 446.1859.

Compounds G ₄ ~ G ₄₈ can be prepared by the same method as the preparation of G ₁ or G ₂ , the difference is:

Adopt BOP, or DCC, or PyBOP, or DIC, or TBTU, or HBTU, or HATU, or TATU to replace EDCI in embodiment 1;

Adopt HOOBt, or HOSu to replace HOBt in embodiment 1;

Adopt 2-(4-(chloromethoxy) phenyl) methyl nitrate or 2-(4-hydroxyl phenyl) acetonitrile or 2-(4-hydroxyl phenyl) methyl nitrate to replace in embodiment 1 2-(4-(chloromethoxy)phenyl)acetonitrile;

Using (S)-2-(tert-butoxycarbonyl)aminobutyric acid, or (S)-2-((tert-butoxycarbonyl)methylamino)propionic acid, or (S)-2-((tert-butoxy Carbonyl)methylamino)butyric acid was substituted for N-tert-butoxycarbonyl-L-alanine in Example 1.

Adopt pyridine, or DBU, or N-methylmorpholine to replace DIEA and triethylamine in Example 1.

In order to better illustrate the effect of the present invention, the experimental proof example of the present invention is given below in conjunction with:

1. Experimental example 1

X-linked inhibitor of apoptosis protein (XIAP) is an important member of the IAP protein family. XIAP inhibits the activity of caspases by directly binding to caspase-3, caspase-7, and caspase-9, thereby inhibiting cell apoptosis. In the present invention, taking G ₁ , G ₂ and G ₃ as examples, the abilities of G ₁ , G ₂ and G ₃ to antagonize the activity of the BIR3 domain of X-linked inhibitor of apoptosis protein (XIAP) are respectively determined in the fluorescence polarization test.

1.1 Experimental materials and instruments

(1) Fluorescently labeled peptide ligands

A colleague synthesized the heptapeptide molecule H-AVPFAQK-(6-FAM)-NH ₂ , and made a fluorescent label on the ε amino group of its lysine branched chain. The structure of the fluorescent peptide was confirmed by ESI-MS, and its purity was higher than 98% by HPLC analysis. The fluorescent peptide was dissolved in ultrapure water, and its solution concentration was determined to be 467 μM by ultraviolet absorption method. The solution was divided into 10 μL portions, wrapped in tin foil to avoid light, and stored at -20°C.

(2) XIAP-BIR3 recombinant protein

XIAP-BIR3 recombinant protein was purchased from R&D Systems Inc. The protein is of E. coli origin and contains residues Asn252-Thr356 with an N-terminal Met and 6-His tag. Each 50μg aliquot is stored in 50μL buffer solution containing 50mM Tris-HCl, 300mM KCl, 50μM zinc acetate, and 1mM DTT at -20°C with a pH value of 8.0.

(3) Reaction buffer

The reaction buffer used in this test contains 20mM Hepes, 2mM dithiothreitol (DTT), 100mM NaCl, 0.1% Bovine serum albumin (BSA), and the pH value is adjusted to 7.4 by a pH meter. All reagents used were purchased from Sigma-Aldrich Company with the highest level of purity.

(4) Preparation of sample solution

Firstly, compounds G ₁ , G ₂ and G ₃ were respectively dissolved in ultrapure water to prepare a 10 mM mother solution, and then the mother solution was diluted to the desired concentration.

(5) Experimental equipment

Perkin Elmer EnVision Multilabel Microplate Reader (EnVision Multilabel Reader), pH meter, UV absorbance detector, 384-well detection plate.

1.2 Experimental method

(1) Prepare 384-well detection plate

First, 2.5nM fluorescent peptide and 250nM XIAP-BIR3 protein were added to the reaction buffer to form a test buffer solution, and incubated for half an hour. At the same time, the sample stocks were diluted into a series of concentrations ranging from 5mM to 5nM.

Then, on a 384-well assay plate, three replicates of each concentration were spotted. First, add 16 μL of the test buffer solution to the wells of the 384-well assay plate with a pipette gun, and then add 4 μL of the sample solution. After mixing, the volume of the solution in the wells becomes 20 μL, so the concentrations of the fluorescent peptide and XIAP-BIR3 become the optimal test concentrations, namely 2 nM and 200 nM, respectively. The final concentration of the samples was varied from 1 mM to 1 nM. Incubate the 384-well assay plate at room temperature for 20-30 minutes, and read with a microplate reader. The assay plate was then incubated for another 20-30 minutes and read again.

(2) Data processing

All data were analyzed using the nonlinear regression curve fitting program of GraphPad Prism.

1.3 Results and discussion

The final test concentrations of the fluorescent peptide and XIAP-BIR3 protein were fixed at 2nM and 200nM, respectively, and the final concentration of the tested samples ranged from 1mM to 1nM. The competitive binding inhibition curve between compound _G1 and XIAP-BIR3 is shown in Figure 4, with mP as the Y axis and log C as the X axis (C is the concentration of the test sample). The results showed that compound G ₁ had better binding affinity to XIAP-BIR3. Due to the competitive binding inhibition of compound _G1 on XIAP-BIR3, its binding curve is S-shaped, that is, the mP value decreases with the increase of the test sample concentration, and when the sample concentration is low enough, the mP value reaches the maximum and does not change anymore. Conversely, when the sample concentration is high enough, the mP value reaches a minimum value, suppressing saturation. The measured data were analyzed by GraphPad Prism, and the IC ₅₀ value of compound G ₁ was 3.6 μM.

Under the same conditions and using the same detection method, the measured IC ₅₀ value of G ₂ was 6.7 μM, and the IC ₅₀ value of G _,3 was 8.3 μM. In addition, the IC ₅₀ value of the positive control substance—tetrapeptide molecule AVPI (Ala-Val-Pro-Ile) was 0.8 μM. Although the IC ₅₀ values of G ₁ , G ₂ and G ₃ are not as good as those of AVPI, compounds G ₁ , G ₂ and G ₃ have more obvious non-peptide characteristics and are expected to act on multiple anticancer targets. The results of fluorescence polarization test showed that compounds G ₁ , G ₂ and G ₃ are new and more efficient non-peptide antagonists of XIAP.

2. Experimental example 2

Taking G1 as _an example, in the MDA-MB-231 breast cancer cell line, the synergistic effect of _G1 and another anticancer drug TRAIL on cell apoptosis was determined.

2.1 Experimental materials and methods

(1) Cell culture and reagents

All compounds were dissolved in DMSO and prepared as stock solutions at a concentration of 10 mM. Cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/mL), streptomycin (100 mg/mL). TRAIL was purchased from PeproTech Inc.

(2) 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) test

Cell viability was tested by MTT assay. First, the cells were placed in a 96-well plate (1×10 ⁶ cells/well), and then the compounds to be tested and TRAIL at corresponding concentrations were added, and maintained in the culture dish for 24, 48, and 96 hours. Finally, 20 μL of MTT (5 mg/mL phosphate buffer) was added to the cell culture dish and incubated at 37° C. for 2 hours. The resulting crystals were dissolved in 200 μL of DMSO, and the absorption intensity was measured at a wavelength of 490 nm with Vector3.

2.2 Results and discussion

First, in the MDA-MB-231 cell line, using the MTT assay, tested the effects of TRAIL at three concentrations of 10ng/mL, 50ng/mL, and 100ng/mL on cell apoptosis when used alone. The results are shown in Figure 5, with the increase of time and the increase of TRAIL concentration, the cell viability decreased. However, in general, even at a concentration of 100ng/mL, the effect of TRAIL on the apoptosis of MDA-MB-231 cancer cells is not too obvious. After 96 hours of cell culture, 65% of cancer cells still survived .

Then, using the MTT assay, we tested the effects of 5μM G ₁ and TRAIL at three concentrations of 10ng/mL, 50ng/mL and 100ng/mL on apoptosis. The results are shown in Figure 5, after 48 hours of cell culture, compound _G1 and TRAIL showed a good dose-dependent synergistic effect, and this synergistic effect was more obvious at higher concentrations. From the results obtained, it can be deduced that the more active the apoptosis, the more pronounced this synergistic effect. For example, after 96 hours of incubation, 100ng/mL TRAIL was used alone, and 65% of the cells survived. However, when TRAIL was used together with 5μM _G1 , only 31% of the cells survived, and the anticancer activity was greatly improved.

The test results show that this type of hybrid drug can effectively antagonize the effect of IAPs, and in the MDA-MB-231 breast cancer cell line, it can make the MDA-MB-231 cells that are resistant to TRAIL become sensitive to TRAIL , thereby accelerating cell apoptosis. This type of hybrid drug can be used as a lead compound of a new anticancer drug in the development and research of a new anticancer drug.

List of Acronyms (in alphabetical order)

The above-mentioned embodiment is a preferred embodiment of the present invention, but the embodiment of the present invention is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplifications should be equivalent replacement methods, and all are included in the protection scope of the present invention.

Claims (10)

1. a hydridization cancer therapy drug, it is characterised in that: described hydridization cancer therapy drug structural formula is as shown in formula I:

Wherein:

X is selected from-SCH₂-,-CH=CH-or-(CH₂)₂One in-substituent group:

Y is selected from-O-or-OCH₂One in O-；

R₁It is selected fromOne in substituent group；

R₂Selected from-CN or-ONO₂In one.

2. the preparation method of the hydridization cancer therapy drug described in claim 1, it is characterised in that: comprise the steps:

1) compound A and inorganic base being dissolved in polar organic solvent, suspension stirs 1～30 minute, is subsequently adding iodomethane Or iodoethane, mixed liquor, under the protection of noble gas, is stirred at room temperature 4～12 hours, concentrates, and gained residue is dissolved in nonpolar In organic solvent, organic phase washed with water, desiccant dryness, concentrate the thick product of gained and cross column purification, obtain colorless oil B；

Wherein, the mol ratio of described compound A, inorganic base, iodomethane or iodoethane is: 1:1～10:1～5；

2) at-10～15 DEG C, the chlorohydrocarbon solution of TFA is added to the chlorohydrocarbon solution of compound B, mixed liquor stirring 0.3 ～5 hours, it is concentrated to give thick product amine C, at-10～15 DEG C, compound C is dissolved in dry chlorohydrocarbon solution, add uncle Aminoacid, condensing agent and the additive of butoxy carbonyl protection, is subsequently adding organic base regulation reactant liquor pH to alkalescence, mixing liquid chamber Temperature is stirred overnight, and reactant liquor with dilute acid soln, dilute alkaline soln, saturated aqueous common salt washing, is dried successively, and concentrating under reduced pressure gained slightly produces Product cross column purification, obtain colorless oil D；

Wherein, the aminoacid of described compound C, tertbutyloxycarbonyl protection, condensing agent, the mol ratio of additive are 1:1～5:1 ～20:0.25～20；

3) being dissolved in by compound D in the mixed solvent of polar organic solvent and water, add highly basic, it is little that suspension is stirred at room temperature 2～7 Time, concentrate, residue diluted with water, with dilute hydrochloric acid solution regulation pH value to acid, aqueous solution non-polar organic solvent extracts Three times, merge organic facies, desiccant dryness, be concentrated to give colorless oil E；

Wherein, described compound D, the mol ratio of highly basic are 1:1～5；

4) at-10～15 DEG C, compound E is dissolved in dry chlorohydrocarbon solution, adds organic base or condensing agent and organic base Mixing chlorohydrocarbon solution, is subsequently adding 2-(4-(chloromethoxy) phenyl) acetonitrile or 2-(4-(chloromethoxy) phenyl) methyl Nitrate or 2-(4-hydroxy phenyl) acetonitrile or 2-(4-hydroxy phenyl) methyl nitrate, mixed liquor under inert gas shielding, Stirred overnight at room temperature, is concentrated to give thick product F ,-10～15 DEG C, is added to the chlorohydrocarbon solution of compound F by TFA, mixing Liquid stirs 0.1～5 hour, concentrates, efficient liquid phase purification, lyophilization, obtains white finished product G；

Wherein, described compound E, 2-(4-(chloromethoxy) phenyl) acetonitrile or 2-(4-(chloromethoxy) phenyl) methyl Nitrate or 2-(4-hydroxy phenyl) acetonitrile or 2-(4-hydroxy phenyl) methyl nitrate, organic base, the mol ratio of condensing agent are 1:1～5:1～20:1～20；

The general structure of described compound A is:Wherein, X is-SCH₂-,-CH=CH-or- (CH₂)₂One in-.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described inorganic base is K₂CO₃、Na₂CO₃、KHCO₃、NaHCO₃, one or more in KOH, NaOH or LiOH；

Described highly basic is KOH, NaOH, Ca (OH)₂, one or more in NaH, KH or LiOH.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described polar organic solvent For one or more in DMF, DMSO, THF or MeCN；

Described non-polar organic solvent is one or more in ethyl acetate, dichloromethane, chloroform or ether.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described noble gas is nitrogen Gas or argon；

Described chlorohydrocarbon solution is dichloromethane or chloroform.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described tertbutyloxycarbonyl is protected The aminoacid protected is N-tert-butoxycarbonyl-l-alanine, (S)-2-(tertbutyloxycarbonyl) aminobutyric acid, (S)-2-((tertiary butyloxycarbonyl Base) methylamino) one or more in propanoic acid or (S)-2-((tertbutyloxycarbonyl) methylamino) butanoic acid.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described condensing agent be BOP, One or more in DCC, EDCI, DIC, PyBOP, TBTU, HBTU, HATU or TATU.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described additive is One or more in HOBt, HOOBt or HOSu.

The preparation method of hydridization cancer therapy drug the most according to claim 2, it is characterised in that: described organic base is three second One or more in amine, pyridine, DBU, DIEA or N-methylmorpholine；

Described desiccant is the one in anhydrous magnesium sulfate, anhydrous sodium sulfate, Anhydrous potassium carbonate or anhydrous calcium chloride.

10. the hydridization cancer therapy drug described in claim 1 is as the application of cancer therapy drug.