CN110862403A - Camptothecin-glycolic acid-norcantharidin conjugate and application thereof - Google Patents
Camptothecin-glycolic acid-norcantharidin conjugate and application thereof Download PDFInfo
- Publication number
- CN110862403A CN110862403A CN201810980917.0A CN201810980917A CN110862403A CN 110862403 A CN110862403 A CN 110862403A CN 201810980917 A CN201810980917 A CN 201810980917A CN 110862403 A CN110862403 A CN 110862403A
- Authority
- CN
- China
- Prior art keywords
- camptothecin
- norcantharidin
- conjugate
- reaction
- glycolic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
The invention provides a camptothecin-glycolic acid-norcantharidin conjugate I:
Description
Technical Field
The invention belongs to the field of new drug design and synthesis, and particularly relates to a novel camptothecin-glycolic acid-norcantharidin conjugate and anti-tumor application thereof.
Background
Camptothecin is an anticancer quinoline alkaloid discovered by Monroe Ewald and Mansukh Canal in 1958, has wide anticancer activity, and is mainly used for treating various cancers such as colon cancer, ovarian cancer, liver cancer, bone cancer, leukemia and the like. Camptothecin is bound to DNA topoisomerase I through its lactone structure to achieve antitumor effect. Norcantharidin (NCTD) is a compound obtained by removing 2, 3-methyl in cantharidin, and is a novel anti-cancer drug which is synthesized artificially preferentially in China. Has low toxicity and also has leukocyte increasing effect. Studies have shown that NCTD can protect liver cells from damage caused by Lipopolysaccharide (LPS); protection of renal tubular fibrosis; also has good immunoregulation function for human body. Therefore, the structural modification using norcantharidin as a primer is the research direction of the subject.
With the increasing mortality and morbidity of cancer, it has become one of the most important problems in our country. Cytotoxic drugs are still the main body of the current tumor treatment drugs, because the cytotoxic drugs act on tumor cells and ordinary cells, the curative effect on solid tumors is poor, the side effect is great, and the drugs have drug resistance, and in consideration of the disease situation of tumors in China, the research and development of efficient, low-toxicity and drug-free tumor drugs are very important. Natural drugs and derivatives thereof are one of the important sources of antitumor drugs, and the natural drugs are still the best means for finding clinical candidate compounds through structural modification. Currently, camptothecin is one of the most widely studied natural products, and three camptothecin compounds have been approved for clinical treatment of some malignancies (irinotecan, topotecan, and belotecan). In addition, the defects of poor water solubility and instability in human physiological environment of the camptothecin still exist, and the immune function of the human body is damaged when the camptothecin is clinically used. The norcantharidin has the characteristics of immunoregulation and leukocytosis. The research on the structure of camptothecin shows that the group necessary for antitumor activity is a lactone ring on an E ring, but an internal alicyclic ring is easy to hydrolyze and loses activity, and the esterification reaction of a hydroxyl group at the 20-position of camptothecin and a molecule with a carboxylic acid group is a common method for stabilizing the lactone ring, because the camptothecin with the lactone structure can be disconnected under physiological conditions and released.
Therefore, the subject uses the camptothecin as a synthetic raw material, uses glycolic acid as a binding intermediate, links camptothecin and norcantharidin derivatives to develop a camptothecin-20 site ester derivative with novel anti-tumor efficacy, and performs activity determination and screening on the camptothecin-20 site ester derivative. Therefore, the purpose that one medicine treats multiple diseases and one medicine has multiple effects is achieved, the camptothecin has good anti-tumor efficacy, the effects of increasing the leucocytes and improving the immunity are achieved, and the development prospect and the application approach are good.
In order to search for anticancer drug candidates with better drug effect, the invention designs a camptothecin-glycolic acid-norcantharidin conjugate with a unique structure, which connects the 20-hydroxyl of camptothecin and norcantharidin together through glycolic acid, and designs a synthesis method to prepare the target derivative with high yield.
Disclosure of Invention
The invention provides a novel camptothecin-glycolic acid-norcantharidin conjugate I:
in formula I, the dotted line on the ring represents that the bond at that position is a saturated or unsaturated double bond; r1 is selected from C1-C6 alkyl, substituted alkyl, cycloalkyl, benzyl or substituted benzyl; r2 is selected from hydrogen or halogen.
In a preferred embodiment, R1 of formula I is selected from R of formula I is selected from C1-C4 alkyl, cycloalkyl or benzyl; more preferably, R1 of formula I is methyl, ethyl, propyl, butyl, cyclopropyl or benzyl.
In a preferred embodiment, the halogen in formula I is selected from fluorine, chlorine or bromine; bromine is preferred.
In another aspect, the present invention provides a method for synthesizing camptothecin-glycolic acid-norcantharidin conjugate I, comprising the following steps: 1) camptothecin and benzyloxy acetic acid are subjected to esterification reaction in the presence of a coupling agent and organic base to obtain a compound II; 2) removing benzyl from the compound II through catalytic hydrogenation reaction to obtain a compound III; 3) and norcantharidin monoester IV are subjected to esterification reaction in the presence of a coupling agent and organic alkali to obtain a camptothecin-glycolic acid-norcantharidin conjugate I, and the synthetic route is as follows:
wherein, R in formula I and formula IV in the synthetic route1And R2The selected groups are kept consistent.
In a preferred embodiment, the coupling agent in step 1) is selected from EDCI, DCC or DIC; the organic base in the step 1) is selected from triethylamine, diisopropylamine, DMAP or DABCO; the solvent is selected from dichloromethane or chloroform.
In a preferred embodiment, the catalyst for the catalytic hydrogenation in step 2) is selected from the group consisting of Pd/C, Pd (OH)2and/C or Pt/C.
In a preferred embodiment, the coupling agent in step 3) is selected from EDCI, DCC or DIC; the organic base in the step 1) is selected from triethylamine, diisopropylamine, DMAP or DABCO; the solvent is selected from dichloromethane or chloroform.
In the above synthetic route, the reaction solvent can be selected from N, N-dimethylformamide, dimethylsulfoxide, dichloromethane, chloroform, tetrahydrofuran or isopropyl ether according to the requirements of the reaction on temperature and polarity of the solvent.
The reaction temperature may be appropriately selected depending on the type of the reaction.
The reaction time can be obtained by tracking the reaction condition through monitoring means such as thin layer chromatography TLC, high performance liquid chromatography HPLC or LC-MS liquid mass spectrum combination and the like.
Common chemical or solvent abbreviations in the present application include: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (abbreviated EDCI), N-dicyclohexylcarbodiimide (abbreviated DCC), N-diisopropylcarbodiimide (abbreviated DIC), 4-dimethylaminopyridine (abbreviated DMAP), 1, 4-diazabicyclo [2.2.2] octane (abbreviated DABCO), N-dimethylformamide (abbreviated DMF), dimethyl sulfoxide (abbreviated DMSO), dichloromethane (abbreviated DCM) and the like.
The activity test proves that the camptothecin-glycolic acid-norcantharidin conjugate I designed and synthesized by the invention has good anti-liver cancer effect. Therefore, in a third aspect, the invention provides the use of camptothecin-glycolic acid-norcantharidin conjugate I for preparing an anti-tumor medicament; preferably, the application of the compound in preparing medicines for resisting liver cancer, gastric cancer, colon cancer and pancreatic cancer.
The invention has the advantages that: the invention provides a camptothecin-glycolic acid-norcantharidin conjugate I, which integrates two active drug fragments of camptothecin and norcantharidin derivatives and is a novel double-target tumor inhibitor. The camptothecin-glycolic acid-norcantharidin conjugate is proved to have good anti-tumor effect by activity tests, and particularly has high activity on liver cancer, gastric cancer, colon cancer and pancreatic cancer. In addition, the method for preparing the camptothecin-glycolic acid-norcantharidin conjugate I has the advantages of easily available raw materials, low cost and high yield of a target product of the synthetic reaction; is easy to prepare.
Detailed Description
The invention will be further illustrated by the following specific examples, which are not intended to limit the scope of the invention. Without departing from the inventive concept, a person skilled in the art may make modifications or combinations of the parameters or conditions of the claims, which modifications or combinations shall also be considered as the protective scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims. The solvent and reagent used in the present invention are from Shanghai reagent company, national drug group. The reagents used are all chemically pure, unless otherwise specified.
EXAMPLE 1 preparation of camptothecin-hydroxyacetic acid intermediate III
(1) Preparation of Compound II
Camptothecin (1g,3.45mmol) and CH2Cl2(35mL) were put in a round bottom flask, stirred at room temperature to give a yellow suspension, DMAP (500mg,4.1mmol,1.2eq), benzyloxyacetic acid (1mL,6.02mmol,1.74eq) and DIPC (5mL,39.68mmol,13eq) were slowly added in this order, and the mixture was continued at normal temperatureThe reaction was stirred at room temperature. Follow the TLC plate and observe the developing reagent (CH2Cl2: CH3OH ═ 97:3) under UV lamp 254 nm. After 12h of reaction, the reaction was stopped, filtered and the filter cake was dried to give the desired product II (1.08g, 100% yield) as a yellow solid, Rf＝0.6(CH2Cl2:CH3OH 97: 3). The product was directly taken to the next reaction without further purification.
(2) Preparation of Compound III
Putting the reactant II (200mg,0.4m mol) into a round-bottomed bottle, adding CH3OH 10ml, and then adding 10% Pd/C (50mg,0.5 mmol); after evacuation of the reaction flask, H2 was passed through and the reaction 24 was stirred at 30 ℃ at room temperature, the TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was observed under an ultraviolet lamp at 254 nm. The reaction was stopped, the catalyst was removed by filtration, the filtrate was spin-dried and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give product III as a yellow solid (116mg, 72% yield). M.p.159-160 ℃ C, Rf＝0.3(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.69(s,1H),8.14(dd,J＝8Hz,8Hz,2H),7.85(t,J＝8Hz,1H),7.70(t,J＝8Hz,1H),7.08(s,1H),5.49(s,1H),5.29(s,1H),4.33(d,J＝16Hz,1H),4.16(d,J＝16Hz,1H),3.42(q,J＝8Hz,2H),2.07-2.17(m,2H),0.95(dt,J＝8Hz,8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝172.17,167.69,157.19,156.98,152.77,148.29,146.41,145.81,132.06,130.89,130.27,129.33,128.99,128.77,128.76,128.43,128.25,128.24,128.21,128.17,119.26,95.43,76.29,66.65,59.78,50.70,30.56,23.73,7.99.
Example 2 preparation of intermediate IV
According to the synthetic method disclosed in the paper (titled "synthesis of camptothecin norcantharidin ester and preliminary evaluation of antitumor activity in vitro", china new medicine journal, 2018, 27(8):99-107), intermediate IV can be easily synthesized: reacting maleic anhydride with furan in ether, and stereoselectively obtaining 5-alkene norcantharidin 1 through [4+2] cycloaddition reaction; the compound 1 is hydrogenated under the catalytic action of Pd/C to obtain norcantharidin 2; meanwhile, the compound 1 is added with liquid bromine under the condition of chloroform to obtain 5, 6-dibromo-norcantharidin 3 with higher yield; the side chain IVa-i of the corresponding 5-alkene norcantharidin monoacid monoester is obtained by alcoholysis of the compounds 1, 2 and 3 under the catalysis of triethylamine base with higher yield (refer to the following synthetic route).
Example 3 preparation of camptothecin-hydroxyacetic acid-norcantharidin conjugate I
(1) Preparation of Compound Ia
The reaction mixture was stirred in a 50mL oil bath at 50 ℃ with reaction mixture III (100mg,0.25mmol), reaction mixture IVa (200mg,1.01mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15 mL). The reaction was followed by TLC plates and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was observed under uv lamp 254 nm. The reaction was stopped for 6h, the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel, the organic phase was left, dried for 15min, filtered, the filtrate was dried, the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to yield yellow solid product Ia (108mg, 74%) m.p.128-129 ℃, r.p.f＝0.41(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.66(s,1H),8.11(dd,J＝8Hz,8Hz,2H),7.83(t,J＝8Hz,1H),7.68(t,J＝8Hz,1H),7.15(d,J＝8Hz,1H),6.42(s,2H),5.26(s,1H),5.09(d,J＝12Hz,1H),5.03(dd,J＝4Hz,4Hz,1H),4.90(s,1H),3.56-3.64(m,4H),3.44(s,3H),2.84(dd,J＝8Hz,8Hz,2H),2.15(q,J＝8Hz,2H),0.92(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝171.87,171.84,171.04,171.03,167.23,167.18,157.20,156.91,152.75,148.31,145.21,136.80,131.89,130.83,130.21,129.32,128.95,128.40,128.12,119.34,96.21,80.50,80.20,77.20,77.17,66.74,52.13,52.00,50.67,46.68,46.34,23.68,7.93.IR(KBr)ν(cm-1)＝3442,2924,2853,1750,1720,1668,1618,1457,1401,1380,1221,1175,838,764,732.HRMS(ESI-TOF):587.1690[M+1].
(2) Preparation of Compound Ib
The reaction mixture was taken out of reaction III (100mg,0.25mmol), reaction IVb (200mg,0.94mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and stirred in a 50 ℃ oil bath for 6 hours. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. After the reaction was stopped, the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel, leaving the organic phase, dried for 15min, filtered, the filtrate was spin-dried, and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give product Ib (120mg, 81%) as a yellow solid. M.p.135-136 deg.C, Rf＝0.52(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.67(s,1H),8.12(dd,J＝8Hz,8Hz,2H),7.84(t,J＝8Hz,1H),7.69(t,J＝8Hz,1H),7.16(d,J＝8Hz,1H),6.41(d,J＝8Hz,1H),5.27(s,1H),5.29(s,1H),5.05(dd,J＝20Hz,8Hz,2H),4.81(dd,J＝20Hz,20Hz,1H),3.81-4.02(m,2H),3.56-3.64(m,4H),2.83(dd,J＝8Hz,8Hz,2H),2.14(q,J＝8Hz,2H),0.90(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝171.30,171.00,167.24,157.21,156.92,152.79,148.32,146.52,145.21,137.24,136.79,132.04,130.85,130.18,129.27,128.97,128.40,128.13,119.21,95.63,80.23,77.29,66.70,60.68,50.70,46.22,30.64,23.72,14.17,7.93.IR(KBr)ν(cm-1)＝3441,3060,2982,2940,1754,1668,1620,1561,1501,1457,1403,1232,1157,1049,811,784.HRMS(ESI-TOF):601.1847[M+1].
(3) Preparation of Compound Ic
The reaction mixture was taken out of reaction III (100mg,0.25mmol), reaction IVc (200mg,0.73mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and stirred in a 50 ℃ oil bath for 6 hours. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. After the reaction was stopped, the reaction solution was washed with aqueous NaHCO3 solution (7mL) in a separatory funnel, leaving the organic phase, dried for 15min, filtered with suction, the filtrate was spin-dried, and the crude product was obtainedColumn chromatography with eluent (CH2Cl2: CH3OH ═ 97:3) afforded the product Ic (114mg, 70%) as a yellow solid. M.p.125-126 ℃, Rf＝0.51(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.58(s,1H),8.10(dd,J＝8Hz,8Hz,2H),7.82(t,J＝8Hz,1H),7.66(t,J＝8Hz,1H),7.19-7.31(m,4H),7.14(d,J＝8Hz,2H),7.02(d,J＝2Hz,1H),6.41(s,1H),4.93-5.21(m,4H),4.75(dd,J＝20Hz,12Hz,2H),3.60(q,J＝8Hz,2H),2.88(dd,J＝8Hz,8Hz,1H),2.80(d,J＝8Hz,1H),0.91(d,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝171.34,171.29,171.19,170.93,170.87,167.35,167.12,157.22,155.88,152.60,148.29,146.40,154.31,130.81,128.77,128.68,128.54,128.50,119.19,95.59,80.52,80.30,77.24,66.52,66.44,60.68,45.42,41.09,40.58,40.16,39.74,39.53,30.57,7.95.IR(KBr)ν(cm-1)＝3432,2921,2854,1753,1663,1618,1560,1402,1154,1058,997,760,723,699.HRMS(ESI-TOF):663.2001[M+1].
(4) Preparation of Compound Id
The reaction mixture was taken out of reaction III (100mg,0.25mmol), reaction IVd (200mg,1.00mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and stirred in a 50 ℃ oil bath for 6 hours. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. After the reaction was stopped, the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel, leaving an organic phase, dried for 15min, filtered, the filtrate was spin-dried, and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give the product Id as a yellow solid (120mg, 83%). M.p.197-198 deg.C, Rf＝0.5(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.36(s,1H),8.20(d,J＝8Hz,1H),7.90(d,J＝8Hz,1H),7.80(t,J＝8Hz,1H),7.63(t,J＝8Hz,1H),7.23(d,J＝2Hz,1H),5.64(dd,J＝20Hz,4Hz,1H),5.37(d,J＝20Hz,1H),5.24(s,1H),4.85-4.90(m,1H),4.93(d,J＝8Hz,1H),4.72(dd,J＝16Hz,16Hz,1H),4.16(d,J＝2Hz,2H),3.81(dd,J＝8Hz,8Hz,1H),3.51-3.60(s,3H),3.00(dd,J＝12Hz,4Hz,2H),2.93(d,J＝8Hz,2H),2.28(dd,J＝8Hz,8Hz,1H),2.16(dd,J＝8Hz,8Hz,1H),1.69-1.77(m,2H),1.45(dd,J＝8Hz,8Hz,2H),0.95(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝171.26,171.23,170.26,170.21,166.97,166.84,166.75,166.69,157.22,156.99,152.19,152.13,148.80,148.77,146.49,146.40,145.08,144.91,131.19,130.67,130.63,129.62,128.38,128.33,128.14,128.12,128.04,120.20,96.04,78.52,78.44,78.42,78.37,76.86,76.82,67.20,67.12,60.65,60.39,52.17,52.04,51.68,51.45,49.94,42.08,31.80,31.76,29.67,29.00,28.98,28.94,28.85,23.47,22.67,14.11,7.50.IR(KBr)ν(cm-1)＝3442,2952,2923,2853,1753,1712,1666,1619,1560,1501,1456,1402,1297,1230,1150,1057,1003,816,787,723.HRMS(ESI-TOF):589.1848[M+1].
(5) Preparation of Compound Ie
The reaction mixture was stirred in a 50mL oil bath for 6h, taking reaction III (0.25mmol), reaction IVe (200mg,0.93mmol), EDCI (100mg,1.04mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and placing them in a 50mL sealed tube. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. The reaction was stopped and the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel leaving an organic phase, dried for 15min, filtered, the filtrate was spin dried and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to yield the yellow solid product Ie (121mg, 82%). M.p.141-143 ℃ and Rf＝0.52(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.66(s,1H),8.14(d,J＝8Hz,1H),8.09(d,J＝8Hz,1H),7.82(t,J＝8Hz,1H),7.67(t,J＝8Hz,1H),7.13(t,J＝8Hz,1H),5.27(s,2H),4.93(dd,J＝8Hz,8Hz,1H),4.77(dd,J＝8Hz,8Hz,1H),4.54-4.71(m,2H),3.76-4.0(m,2H),3.13(d,J＝8Hz,1H),3.02(d,J＝8Hz,1H),2.08-2.18(m,1H),1.39-1.55(m,4H),1.1(dt,J＝8Hz,8Hz,3H),0.90(t,J＝8Hz,3H),0.8(q,J＝8Hz,2H).13C NMR(100MHz,DMSO-d6)δ＝170.88,170.54,167.16,157.22,156.91,152.71,148.37,146.47,146.27,132.01,131.95,130.80,130.75,129.35,128.94,128.68,128.22,128.15,128.09,128.07,119.30,95.55,79.57,78.40,78.31,78.13,77.12,76.73,72.67,66.71,60.88,60.57,60.48,60.40,52.31,51.58,51.48,51.30,50.62,41.08,39.96,39.75,30.69,29.48,28.87,28.75,23.71,22.53,14.43,14.28,14.14,7.96.IR(KBr)ν(cm-1)＝3448,3061,2957,2823,1753,1664,1616,1566,1501,1458,1401,1296,1261,1232,1191,1142,1085,1056,801,724,560.HRMS(ESI-TOF):603.1986[M+1].
(6) Preparation of Compound If
The reaction mixture was taken out of reaction III (100mg,0.25mmol), reaction IVf (200mg,0.72mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and stirred in a 50 ℃ oil bath for 6 hours. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. After the reaction was stopped, the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel, leaving an organic phase, dried for 15min, filtered, the filtrate was spin-dried, and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give yellow solid product If (145mg, 89%). M.p.127-129 ℃, Rf＝0.53(DCM:CH3OH＝97:3).1H NMR(400MHz,CDCl3)δ＝8.23(s,1H),7.90(dd,J＝8Hz,8Hz,1H),7.80(t,J＝8Hz,1H),7.64(t,J＝8Hz,1H),7.21-7.31(m,5H),7.15(t,J＝8Hz,1H),6.94(d,J＝2Hz,1H),5.66(t,J＝16Hz,1H),5.26(dd,J＝4Hz,4Hz,1H),5.03(s,1H),4.95(s,1H),4.89(d,J＝8Hz,1H),4.87(d,J＝8Hz,2H),4.72(d,J＝4Hz,1H),4.65(d,J＝4Hz,1H),3.59(t,J＝8Hz,1H),3.08(d,J＝8Hz,1H),2.96(d,J＝8Hz,1H),2.1-2.31(m,2H),1.65-1.81(m,4H),0.98(q,J＝8Hz,3H).13C NMR(100MHz,CDCl3)δ＝174.70,172.15,170.80,170.63,170.15,166.93,166.88,166.80,166.75,157.32,157.25,152.10,151.86,148.73,148.67,146.43,146.32,145.81,145.23,135.54,135.50,135.15,131.28,131.05,130.76,130.72,129.62,129.58,128.66,128.59,128.55,128.51,128.49,128.39,128.36,128.34,128.28,128.26,128.20,128.17,128.15,128.10,129.09,128.06,120.15,119.66,96.41,96.33,80.70,78.49,77.85,67.12,67.06,66.90,66.84,66.71,60.53,60.18,52.13,51.83,51.68,51.37,51.06,50.90,50.07,49.94,31.80,31.58,29.68,29.19,28.95,28.85,25.77,22.68,14.12,7.56,7.50.IR(KBr)ν(cm-1)＝3445,2954,2882,1752,1661,1616,1499,1455,1385,1351,1295,1175,1054,1001,929,752,699,593.HRMS(ESI-TOF):600.1997[M+1].
(7) Preparation of Compound Ig
The reaction mixture was taken out of reaction III (100mg,0.25mmol), reaction IVg (200mg,0.56mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) and stirred in a 50 ℃ oil bath for 6 hours. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. The reaction was stopped and the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel leaving an organic phase, dried for 15min, filtered, the filtrate was spin dried and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to yield the yellow solid product Ig (138mg, 75%). M.p.137-138 ℃, Rf＝0.42(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.39(s,1H),8.23(dd,J＝8Hz,8Hz,1H),7.93(d,J＝8Hz,1H),7.82(t,J＝8Hz,1H),7.66(t,J＝8Hz,1H),7.33(d,J＝8Hz,1H),5.68(t,J＝16Hz,1H),5.40(dd,J＝16Hz,8Hz,1H),5.28(s,2H),4.81-5.03(m,2H),4.57-4.68(m,2H),4.29(s,3H),3.68-3.75(m,2H),2.14-2.31(m,2H),1.11-1.14(m,2H),0.97(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝176.03,164.39,147.04,143.63,140.24,131.60,126.93,126.83,126.76,126.04,125.52,124.37,123.58,123.37,123.28,123.25,123.22,123.02,122.98,122.96,122.87,122.83,115.11,109.99,90.74,68.12,62.00,61.39,44.77,26.60,24.48,8.90,2.23.IR(KBr)ν(cm-1)＝3446,2936,2854,1753,1664,1617,1565,1500,1455,1437,1402,1285,1232,1165,1056,995,947,899,761,723.
(8) Preparation of Compound Ih
Reaction III (100mg,0.25mmol), reaction IVh (200mg,0.54mmol), EDCI (100mg,0.52mmol), DMAP (20mg,0.16mmol) and CH2Cl2(15mL) were taken in a 50mL sealed tube and stirred in a 50 ℃ oil bath for 6 h. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. After the reaction was stopped, the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel, leaving an organic phase, dried for 15min, filtered, the filtrate was spin-dried, and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give the product Ih as a yellow solid (148mg, 79%). M.p.138-139 ℃, Rf＝0.51(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.63(s,1H),8.10(dd,J＝8Hz,8Hz,2H),7.83(t,J＝8Hz,1H),7.68(t,J＝8Hz,1H),7.14(dd,J＝8Hz,2Hz,1H),5.23(d,J＝4Hz,2H),4.94(t,J＝8Hz,1H),4.83(dd,J＝8Hz,8Hz,1H),4.69(dd,J＝20Hz,4Hz,1H),4.51(dd,J＝8Hz,8Hz,1H),3.98-4.06(m,2H),3.58-3.63(m,2H),3.47-3.53(m,1H),3.22(t,J＝8Hz,1H),2.16(t,J＝8Hz,2H),1.06-1.16(m,3H),0.91(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝171.30,170.49,170.43,169.56,169.17,167.35,167.18,167.07,167.01,157.20,156.88,152.74,152.70,148.35,148.30,146.51,146.43,145.28,145.04,137.73,132.05,132.01,131.98,131.95,131.92,131.91,130.90,130.88,130.84,130.23,130.18,130.15,129.37,129.35,129.33,129.30,128.95,128.76,128.74,128.59,128.47,128.38,128.24,128.20,128.16,128.14,119.39,119.18,95.54,95.50,90.64,88.70,88.62,86.74,77.54,77.32,77.28,76.79,72.67,66.76,66.75,66.72,66.65,66.64,61.88,61.68,60.18,54.34,54.19,54.00,51.96,51.85,50.64,49.12,49.04,48.99,48.65,30.76,30.56,30.45,21.19,14.51,14.34,14.21,14.06,8.00.IR(KBr)ν(cm-1)＝3443,2977,2935,1753,1664,1617,1565,1500,1545,1402,1294,1231,1167,1054,1013,947,888,761.HRMS(ESI-TOF):761.0163[M+1].
(9) Preparation of Compound Ii
Take reactant III (100mg,0.25mmol), reactant Ii (200mg,0.46mmol), EDCI (100mg,0.52mmol)mmol), DMAP (20mg,0.16mmol), CH2Cl2(15mL) were placed in a 50mL sealed tube and the reaction stirred in a 50 ℃ oil bath for 6 h. TLC plates were spotted and the developing reagent (CH2Cl2: CH3OH ═ 97:3) was viewed under UV lamp 254 nm. The reaction was stopped and the reaction was washed with aqueous NaHCO3 (7mL) in a separatory funnel leaving an organic phase which was dried for 15min, filtered, the filtrate was dried and the crude product was purified by column chromatography using an eluent (CH2Cl2: CH3OH ═ 97:3) to give the product Ii (162mg, 80%) as a yellow solid. M.p.123-124 ℃, Rf＝0.53(DCM:CH3OH＝97:3).1H NMR(400MHz,DMSO-d6)δ＝8.33(s,1H),8.21(t,J＝8Hz,1H),7.90(dd,J＝8Hz,8Hz,1H),7.80(t,J＝8Hz,1H),7.64(t,J＝8Hz,1H),7.24-7.29(m,4H),7.16(t,J＝8Hz,1H),6.94(d,J＝8Hz,1H),5.63(t,J＝8Hz,1H),5.34(dd,J＝8Hz,8Hz,1H),5.18(d,J＝4Hz,1H),5.04(d,J＝8Hz,1H),4.95(d,J＝8Hz,1H),4.85(d,J＝8Hz,1H),4.75(d,J＝8Hz,1H),4.70(d,J＝8Hz,1H),4.24(dt,J＝8Hz,1H),3.91(dd,J＝8Hz,8Hz,1H),3.80(q,J＝8Hz,1H),3.04(dd,J＝40Hz,4Hz,1H),2.20-2.29(m,2H),2.09-2.15(m,2H),0.99(t,J＝8Hz,3H).13C NMR(100MHz,DMSO-d6)δ＝169.15,169.14,168.90,168.76,166.91,166.86,166.42,166.38,157.24,157.18,152.13,151.90,148.72,148.66,146.46,146.36,145.60,145.08,140.92,134.99,134.65,131.24,131.02,130.76,130.72,129.59,129.55,128.81,128.64,128.60,128.59,128.50,128.44,128.42,128.36,128.32,128.29,128.18,128.17,128.09,128.06,127.56,126.96,120.04,119.62,96.11,96.03,86.34,86.29,82.48,82.47,77.25,77.15,76.91,67.51,67.37,67.12,66.09,65.24,60.85,60.57,53.60,53.58,53.46,52.64,49.98,49.86,49.68,49.48,46.55,46.24,31.78,31.54,29.69,7.53.IR(KBr)ν(cm-1)＝3451,2939,2884,1753,1664,1617,1562,1500,1455,1385,1349,1231,1165,1056,946,811,756.HRMS(ESI-TOF):823.0324[M+1].
Example 3 solubility test
The compound Ia obtained by the synthesis and the parent compound camptothecin were selected, dissolved in chloroform at 25 ℃ and the solubility results are shown in Table 1.
TABLE 1 solubility of Compounds Ia and camptothecin in chloroform at 25 ℃
Compound Ia has several times the solubility of camptothecin in chloroform. It can be seen that the solubility of the synthetic product I in organic solvents is much better than that of camptothecin.
Example 4 bioactivity test experiment
Cell line and solvent
Tumor cell lines: human liver cancer cell HEPG2, human gastric cancer cell BGC803, human colon cancer cell SW480, human pancreatic cancer cell PANC-1 cell cultured in RPMI1640 containing 10% fetal calf serum
Solvent: dimethylsulfoxide (abbreviated as DMSO).
Embodiment for detecting cell anti-tumor activity by MTT method
In the test, cantharidin is taken as a positive control, DMSO solvent is taken as a blank control, and the inhibitory activity test of camptothecin-glycolic acid-norcantharidin conjugate with the concentration of 50 mu mol/mL on four tumor cells, namely liver cancer cell HEPG2, human gastric cancer cell BGC803, colon cancer cell SW480 and pancreatic cancer PANC-1, is carried out. After 72h of drug treatment, the inhibition using camptothecin-hydroxyacetic acid-norcantharidin conjugate I was evaluated using a standard MTT assay.
TABLE 2 inhibitory Activity of camptothecin-hydroxyacetic acid-norcantharidin conjugate I on four tumor cells
atest solvent is DMSO.
Camptothecin, glycolic acid and functionalized norcantharidin are constructed into conjugates through two coupling reactions, and a series of novel camptothecin-glycolic acid-norcantharidin conjugates I are obtained with better yield. The synthesized conjugate I is tested to have strong inhibitory activity on cancer cells in vitro, in particular to four cancer cell lines of human liver cancer cell HEPG2, gastric cancer cell BGC803, colon cancer cell SW480 and pancreatic cancer PANC-1; can be used for preparing corresponding anti-tumor candidate drugs.
Claims (9)
1. Camptothecin-glycolic acid-norcantharidin conjugate I:
in formula I, the dotted line on the ring represents that the bond at that position is a saturated or unsaturated double bond;
r is selected from C1-C6 alkyl, substituted alkyl, cycloalkyl, benzyl or substituted benzyl; r2 is selected from hydrogen or halogen.
2. The camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to claim 1, wherein R of formula I is selected from C1-C4 alkyl, cycloalkyl or benzyl.
3. The camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to claim 2, wherein R of formula I is selected from methyl, ethyl, propyl, butyl, cyclopropyl or benzyl.
4. The method for synthesizing camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to any one of claims 1 to 3, comprising the following steps:
1) camptothecin and benzyloxy acetic acid are subjected to esterification reaction in the presence of a coupling agent and organic base to obtain a compound II; 2) removing benzyl from the compound II through catalytic hydrogenation reaction to obtain a compound III; 3) and norcantharidin monoester IV are subjected to esterification reaction in the presence of a coupling agent and organic alkali to obtain a camptothecin-glycolic acid-norcantharidin conjugate I, and the synthetic route is as follows:
wherein, in the synthetic routeR in formula I and formula IV1And R2The selected groups are kept consistent.
5. The method for synthesizing camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to claim 4, wherein the coupling agent in step 1) is selected from EDCI, DCC or DIC; the organic base in the step 1) is selected from triethylamine, diisopropylamine, DMAP or DABCO; the solvent is selected from dichloromethane or chloroform.
6. The method for synthesizing camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to claim 4, wherein the catalyst for catalytic hydrogenation reaction in step 2) is selected from Pd/C, Pd (OH)2and/C or Pt/C.
7. The method for synthesizing camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to claim 4, wherein the coupling agent in step 3) is selected from EDCI, DCC or DIC; the organic base in the step 1) is selected from triethylamine, diisopropylamine, DMAP or DABCO; the solvent is selected from dichloromethane or chloroform.
8. The use of camptothecin-hydroxyacetic acid-norcantharidin conjugate I according to any of claims 1 to 3 for the preparation of an antitumor drug.
9. The use according to claim 7, wherein the tumor is selected from liver cancer, stomach cancer, colon cancer or pancreatic cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810980917.0A CN110862403A (en) | 2018-08-27 | 2018-08-27 | Camptothecin-glycolic acid-norcantharidin conjugate and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810980917.0A CN110862403A (en) | 2018-08-27 | 2018-08-27 | Camptothecin-glycolic acid-norcantharidin conjugate and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110862403A true CN110862403A (en) | 2020-03-06 |
Family
ID=69651114
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810980917.0A Pending CN110862403A (en) | 2018-08-27 | 2018-08-27 | Camptothecin-glycolic acid-norcantharidin conjugate and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110862403A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003095460A1 (en) * | 2002-05-06 | 2003-11-20 | The Stehlin Foundation For Cancer Research | Cascade esters of camptothecins and methods of treating cancer using these compounds |
| CN102516347A (en) * | 2011-12-15 | 2012-06-27 | 东北林业大学 | Camptothecin 20-site cholic acid derivative and preparation method thereof |
| CN104817574A (en) * | 2015-05-26 | 2015-08-05 | 遵义医学院 | Novel camptothecin derivative and antitumor application thereof |
| CN105399757A (en) * | 2015-12-29 | 2016-03-16 | 遵义医学院 | Acid-sensitive camptothecin-site 20 norcantharidate derivative and antineoplastic application thereof |
| CN105646546A (en) * | 2015-12-29 | 2016-06-08 | 遵义医学院 | Acid-sensitive camptothecin-20-position ester derivative and antineoplastic application thereof |
-
2018
- 2018-08-27 CN CN201810980917.0A patent/CN110862403A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003095460A1 (en) * | 2002-05-06 | 2003-11-20 | The Stehlin Foundation For Cancer Research | Cascade esters of camptothecins and methods of treating cancer using these compounds |
| CN102516347A (en) * | 2011-12-15 | 2012-06-27 | 东北林业大学 | Camptothecin 20-site cholic acid derivative and preparation method thereof |
| CN104817574A (en) * | 2015-05-26 | 2015-08-05 | 遵义医学院 | Novel camptothecin derivative and antitumor application thereof |
| CN105399757A (en) * | 2015-12-29 | 2016-03-16 | 遵义医学院 | Acid-sensitive camptothecin-site 20 norcantharidate derivative and antineoplastic application thereof |
| CN105646546A (en) * | 2015-12-29 | 2016-06-08 | 遵义医学院 | Acid-sensitive camptothecin-20-position ester derivative and antineoplastic application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 胥佩菱: "《抗癌药物合成设计》", 31 December 1993 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2021513555A5 (en) | Pyrimidine condensed cyclic compound, its production method, and its use | |
| CA2067491A1 (en) | Camptothecin analogs as potent inhibitors of human colorectal cancer | |
| CN113698401B (en) | Beta-elemene macrocyclic derivatives, preparation method and application thereof | |
| CN105399757A (en) | Acid-sensitive camptothecin-site 20 norcantharidate derivative and antineoplastic application thereof | |
| CN109180681B (en) | DNA toxic dimer compound | |
| Nagamura et al. | Synthesis and antitumor activity of duocarmycin derivatives: modification of segment A of duocarmycin B2 | |
| CN111848718A (en) | Tripterine nitrogenous five-membered heterocyclic derivative and preparation method and application thereof | |
| CN111196801A (en) | Aporphine alkaloid derivative and preparation method and application thereof | |
| CN110143974B (en) | Novel artemisinin derivative, synthesis method and application thereof | |
| CN102432622B (en) | 4-amino oxadiazole epipodophyllotoxin derivative and preparation method and application thereof | |
| CN110156817B (en) | Anti-tumor derivative of bi-evodiamine molecule and preparation and application thereof | |
| HU212579B (en) | Process for producing 6-amino-octahydroindolizinetriol derivatives and pharmaceutical compositions containing the same | |
| EP1200444B1 (en) | Novel xanthone compounds, their preparation and use as medicament | |
| KR900006234B1 (en) | Nonel 3',4'-dinitrogen substituted epipodophyllotoxin glucoside derivatiues | |
| CN110862403A (en) | Camptothecin-glycolic acid-norcantharidin conjugate and application thereof | |
| Tsujimoto et al. | Chemistry of ecteinascidins. Part 4: preparation of 2′-N-acyl ecteinascidin 770 derivatives with improved cytotoxicity profiles | |
| Zhao et al. | Synthesis of dual target CPT-Ala-Nor conjugates and their biological activity evaluation | |
| CN112480129B (en) | Polycyclic spiroindoline compound containing guanidyl structural unit and preparation method and application thereof | |
| CN110437265B (en) | Homocamptothecin norcantharidinate derivative and regioselective synthesis method thereof | |
| CN110759961B (en) | Ursolic acid indolyquinone amide derivatives and preparation method and application thereof | |
| EP4019517A1 (en) | 2-aminopyrimidine compounds and pharmaceutical compositions and uses thereof | |
| EP3275882A1 (en) | Process of synthesizing substituted pyridine and pyrimidine compound | |
| US6187783B1 (en) | Phenanthridinium derivatives | |
| EP1377588B1 (en) | 4-beta-1''-[(2''-(substituted benzoyl)anilino] podophyllotoxin analogues useful as anticancer agents | |
| CN110437264B (en) | Homocamptothecin 5, 6-dibromo norcantharidinate derivative and regioselective synthesis method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200306 |