CN106046008A - Chlorin p6 amino acid derivative, preparation method therefor and use of chlorin p6 amino acid derivative - Google Patents

Chlorin p6 amino acid derivative, preparation method therefor and use of chlorin p6 amino acid derivative Download PDF

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CN106046008A
CN106046008A CN201610356755.4A CN201610356755A CN106046008A CN 106046008 A CN106046008 A CN 106046008A CN 201610356755 A CN201610356755 A CN 201610356755A CN 106046008 A CN106046008 A CN 106046008A
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chlorin
amino acid
reaction
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姚建忠
孟志
***
盛春泉
缪震元
董国强
贾宁阳
韩贵焱
刘明辉
刘娜
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Second Military Medical University SMMU
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    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
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    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A61K41/0071—PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines

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Abstract

The invention relates to the technical field of medicines and particularly relates to a novel chlorin photosensitizer, i.e., a chlorin p6 amino acid derivative, a preparation method therefor and use of the chlorin p6 amino acid derivative in preparation of antitumor drugs. The chlorin p6 amino acid derivative disclosed by the invention has a chemical structure represented by a general formula (I) shown in the description. The chlorin p6 amino acid derivative disclosed by the invention has the advantages of high efficiency, low toxicity and the like and can be used for preparing novel photodynamic cancer therapy drugs, drugs for photodynamic therapy on benign vascular diseases such as senile macular degeneration and nevus flammeus and drugs for photodynamic therapy on verruca acuminata.

Description

Chlorin p6 amino acid derivant and its production and use
Technical field
The present invention relates to pharmaceutical technology field, being specifically related to is a kind of novel dihydro porphin photosensitizer chlorin P6 amino acid derivant and preparation method thereof, and the purposes in preparing antitumor drug.
Background technology
Photodynamic therapy (photodynamic therapy, PDT) last century the eighties gradually grow up Neoplasm targeted therapy new technique is i.e. administered (photosensitizer) afterwards by the red light district maximum absorption wave appearance that wavelength is intrinsic with photosensitizer Laser orientation irradiation focus (tumor) tissue joined, by the substrate oxygen (O in photosensitizer induction tissue2) excite generation singlet oxygen (1O2) isoreactivity oxygen species (ROS), cause apoptosis of tumor cells or necrosis to play neoplasm targeted therapy effect.Due to PDT only Tumor tissues is implemented targeting illumination, optionally destroys tumor cell normal tissue or organ then almost without infringement Or infringement is the least, the most also has good controlling, therefore, it is a kind for the treatment of technology non-invasive to human body, Have side effect little (wound produced without operation is painful, the vomiting produced without radiation and chemotherapy, nauseating and immunosuppressant), can be single Solely or coordinate the advantages such as other treatment means repeatedly use.
Oxygen and photosensitizer in illumination, tissue are the three big key elements of PDT, and wherein, photosensitizer is core.First generation porphyrin Photosensitizer such as porfimer sodium (porfimer sodium) etc. is used successfully to clinical therapy of tumor, achieves the curative effect attracted people's attention, But there is also open defect: the laser penetration that (1) red light district maximum absorption wave length (630nm) makes wavelength matched is killed Hinder tumor depth inadequate；And molar absorption coefficient (ε) is little, causes photosensitive activity low；(2) it is multicomponent porphyrin mixture；(3) body It is big that interior removing causes being detained phototoxicity slowly.Therefore, from late nineteen nineties in last century, carried out chlorin such as benzoporphyrin to spread out Biological (benzoporphyrin derivative, BPD) and the second filial generation photosensitizer that chlorophyll a degraded derivant is representative Research and development.Owing to dihydro porphin photosensitizer has single clear and definite, red light district (＞ 600nm) the maximum absorption wavelength relatively porphyrin of structure Red shift causes this wavelength laser to have more preferably the tumor-killing degree of depth and the high number of molar absorption coefficient (ε) to 660-690nm Magnitude, photosensitive activity are strong, internal metabolism is fast, it is little to be detained phototoxicity, it has also become photosensitive new drug research focus.Wherein, Verteporfin (verteporfin), temoporfin (temoporfin) and talaporfin (talaporfin) respectively at 2000, calendar year 2001 and Within 2004, successfully list.
Chlorin p6 extremely unstable, itself 6-with γ-carboxyl can become more stable hexa-atomic inner-acid anhydride to tie by dehydrating condensation voluntarily Structure and generate purpurin 18, therefore, for the structural modification of chlorin p6, document report is less, mainly at its 3-ethylene Base, 13-, 15-carboxyl and three positions of 20-intermediary hydrogen carry out respectively structure of modification (Yao Jianzhong, etc. chlorin p6 ethers spreads out Biological synthesis. organic chemistry, 2001,21 (6): 458-462；Yao Jianzhong, etc. the chlorophyllous degraded of Bombyx mori L. and chlorin p6 The synthesis of amide derivatives. Chinese Journal of Pharmaceuticals, 1999,30 (9): 403-406；Wang Yi, etc. δ-(dilute base of formyl second) two The synthesis of hydrogen porphine derivative. organic chemistry, 2007,27 (3): 391-396；Leach,M.W.；et al.In vitro photodynamic effects of lysyl chlorine p6:cell survival,localization and ultrastructural changes.Photochemistry and Photobiology,1993,58(5):653-660.)。
Its medicine can be improved in view of document report porphyrin and chlorin molecular structure introduce water-soluble amino acid residue Effect and pharmacopedics character (Hitoshi, T., et al.Synthesis of chlorophyll-amino acid conjugates as models for modification of proteins with chromo/ fluorophores.Bioorg.Med.Chem.2014,22,1421-1428.；Kwitniewski,M.,et al.Diamino acid derivatives of PpIX as potential photosensitizers for photodynamic therapy of squamous cell carcinoma and prostate cancer:in vitro studies.J.Photochem.Photobiol.B 2009,94,214–222.；Serra,V.V.,New porphyrin amino acid conjugates:synthesis and photodynamic effect in human epithelial cells.Bioorg.Med.Chem.2010,18,6170–6178.；Wang,H.M.；Porphyrin with amino acid moieties:a tumor photosensitizer.Chem.Biol.Interact.2008,172,154-158.)；It addition, The ether derivative of the 2-position vinyl of chlorin has higher photosensitive activity and tumor PDT curative effect (Yao than its primer Build loyalty etc., the synthesis of chlorin f methyl ether and photosensitization power thereof and tumor photobiological activity. Acta Pharmaceutica Sinica, 2000,35 (1): 63-66；Yao Jianzhong etc., 2-(1-hydroxyl) ethyl chlorin f and the synthesis of ether derivative thereof and tumor photobiological activity. in State's pharmaceutical chemistry magazine, 2001,39 (1): 1-4.), it is photosensitive that the present invention intends researching and developing new a kind of dihydro porphin on this basis Agent.
There is no document report at present using chlorin p6 as guide structure, introduce amino acid residue at its 7-position third carboxyl Or carry out being etherified structure design at its 7-position third carboxyl introducing amino acid residue and 2-position vinyl simultaneously.
Summary of the invention
It is an object of the invention to provide a kind of chlorin p6 amino acid derivant and include that chlorin p6 aminoacid spreads out Biology and chlorin p6 ethers amino acid derivativges.Another object of the present invention is to provide this chlorin p6 amino acid The preparation method of derivant.The third object of the present invention is to provide this chlorin p6 amino acid derivant and is preparing antitumor The particularly purposes in photosensitizer in medicine.
The present invention is with Stability Analysis of Structures and prepares convenient chlorophyll a and stablizes catabolite purpurin 18 as raw material, through interior acid The chlorin p6 dimethyl ester that acid anhydride alkali open ring, carboxyl Azimethylene. methylate and ester selectivity acid hydrolysis prepares is guide structure, Its 7-position third carboxyl introduces water-soluble amino acid residue or introduces after water-soluble amino acid residue at its 7-position third carboxyl while 2-position vinyl is etherified, optimizes design and synthesized a class novel dihydro porphin photosensitizer chlorin p6 class ammonia Base acid derivative.
A first aspect of the present invention, it is provided that a kind of chlorin p6 amino acid derivant and pharmaceutical salts, the present invention The chemical constitution of described chlorin p6 amino acid derivant is as shown in formula I:
In formula I,
R1Represent various amino acid residue；Preferably lysine, glutamic acid, aspartic acid etc..
R2Represent CH=CH2、CH(OR3)CH3；
R3Represent H, low alkyl group, senior alkyl, (CH2)mOR4Or (CH2)mNR5R6；Preferably CH3、C2H5、n-C3H7、n- C4H9、n-C5H11、n-C6H13、n-C8H17Deng.
R4Represent H, low alkyl group；
R5And R6Represent low alkyl group independently；
M represents the integer between 2-6；
Described low alkyl group, refers to the straight or branched alkyl containing 1-6 carbon atom；
Described senior alkyl, refers to the straight or branched alkyl containing 7-18 carbon atom.
Chlorin p6 amino acid derivant of the present invention, part preferred compound I1～I24R1And R2Respectively Combination is as shown in table 1:
Table 1: part preferred compound I of the present invention1～I24
Wherein n represents " n-".
Further, present invention also offers the pharmaceutical salts of above-mentioned chlorin p6 amino acid derivant and pharmaceutically may be used Accepting preparation, described pharmaceutical salts can be alkali metal salt, particular certain cancers etc., described preparation preferred liposome and fat milk etc. Intravenous formulations.
A second aspect of the present invention, it is provided that the preparation method of above-mentioned chlorin p6 amino acid derivant.
The present invention provides the novel dihydro porphin that a kind of raw material sources are wide, preparation method is simple, toxicity is low photosensitive The preparation method of agent chlorin p6 amino acid derivant.
The present invention is raw materials used stablizes catabolite purpurin 18 (purpurin-18) for chlorophyll a, its chemistry knot Structure formula is as follows:
Purpurin 18 is through inner-acid anhydride alkali open ring, carboxyl Azimethylene. methylate and ester selectivity acid hydrolysis prepares dihydro 7-the third carboxyl in porphin p6 dimethyl ester structure and amino acid condensation or its 2-vinyl and 7-the third carboxyl respectively with alcohol and amino Acid occurs addition and condensation reaction, synthesis to obtain the dihydro porphin photosensitizer chlorin of the new structure type of the present invention P6 amino acid derivant.
Above-mentioned purpurin 18 raw material can be prepared through soda acid degraded with the chlorophyll a in Bombyx mori L..
China is silkworm silk big producing country of the world, and Bombyx mori L. (family's silkworm faeces) is one of Main By product of silkworm industry.The annual silkworm of China Silk about can produce 1,000,000 tons of Bombyx mori L.s in producing.Bombyx mori L. Determination of Chlorophyll a content accounts for 0.75%, is extremely to enrich and cheap leaf Verdazulene resource.Utilizing Bombyx mori L. to prepare coke demagnesium chlorophyllin a ethers amino acid derivativges, the medicine that can expand silkworm industry waste resource should With, final realization " turning waste into wealth ".
Purpurin 18 can with literature method synthesis obtain (Yao Jianzhong, etc. the chemistry of Bombyx mori L. Determination of Chlorophyll a degradation process grinds Study carefully. Chinese herbal medicine, 1999,30 (8): 568-57.)
The preparation method of chlorin p6 amino acid derivant of the present invention, with Bombyx mori L. chlorophyll crude extract Commercially available chlorophyll paste is that the preparation method of initiation material is as follows:
A, use chlorophyll a (chlorophyll a, VIII) are that chlorin p6 dimethyl ester prepared by raw material (chlorinp6dimethylester, IV), reaction process is:
B, preparing target compound chlorin p6 amino acid derivatives I by compound III, reaction process is as follows:
Concrete synthesis step is:
A1, prepare purpurin 18 (purpurin-18, VI):
First, the diethyl ether solution of commercially available chlorophyll paste, with the stirring reaction of equal volume amounts concentrated hydrochloric acid under the conditions of 0～5 DEG C 0.5h generates pheophorbide acid a (pheophorbide a, VII)；Then, compound VII is at the normal propyl alcohol liquid of 25% potassium hydroxide In logical oxygen reaction 2 prepare VI；
A2, prepare chlorin p6 dimethyl ester (chlorin p6dimethylester, IV)
Compound VI is dissolved in oxolane-methanol (1:4, v/v), adds equal-volume 0.5mol.L-1 sodium hydroxide reaction 2h system Unstable product chlorin p6 (chlorine p6) afterwards rapidly and diazomethane reaction prepares chlorin p6 trimethyl (chlorin p6trimethylester, V), compound V prepares IV through 25%HCl hydrolysis；
B1, prepare chlorin p6 ether derivative II
The 33%HBr glacial acetic acid liquid room temperature reaction 24h of compounds Ⅳ with excess is prepared intermediate III, and intermediate III is in mistake Amount K2CO3In the presence of with various alcohol (R3OH) reaction prepares compound ii；
B2, prepare chlorin p6 amino acid target derivative I:
By compounds Ⅳ or II dichloromethane solution at 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDCI), in the presence of N-hydroxybenzotriazole (HOBt) and N, N-dimethyl isopropyl amine (DIPEA) respectively with various carboxyls or/ Acidity or hydrochloride of basic amino acid (R with amido protecting1'-NH2HCl) slough through trifluoracetic acid (TFA) again after reaction 24h The tert-butyl group or tertbutyloxycarbonyl or and Lithium hydrate demethylation ester prepare chlorin p6 amino acid target derivative I.
The part preferred compound I of the present invention1～I24R1And R2It is respectively combined table 1 as the aforementioned.
A third aspect of the present invention, it is provided that the pharmaceutical applications of above-mentioned chlorin p6 amino acid derivant.
Chlorin p6 amino acid Derivatives In Mice melanoma cell B16-F10 of the present invention and Humanmachine tumour are thin Born of the same parents A375 etc. has excellent PDT lethal effect, and has efficient, the advantage of low toxicity.
Compared with the photosensitizer Verteporfin (verteporfin) of new generation of existing clinical practice, chlorin of the present invention The advantages such as p6 amino acid derivant has efficiently, low toxicity, can be used for preparing antitumor drug, the newest Photodynamic therapy of cancer Medicine, optical dynamic therapy non-malignant vascular disease such as senile degeneration of macula (a kind of retinal microvascular proliferative disease) and fresh (human papillomavirus feels for the medicine of erythema nevus (a kind of congenital cutaneous blood capillary deformity pathological changes) and optical dynamic therapy condyloma acuminatum Dye disease) medicine.
Detailed description of the invention
In conjunction with embodiment, the present invention is described in detail, but the enforcement of the present invention is not limited only to this.Present invention examination used Agent and raw material the most maybe can be prepared by literature method.
The experimental technique of unreceipted actual conditions in the following example, generally according to normal condition, or according to manufacturer Proposed condition.
Embodiment 1: the preparation of pheophorbide acid a (VII)
Bombyx mori L. chlorophyll a (VIII) crude extract (chlorophyll paste), purchased from Feng Ming chlorophyll company limited of Haining city.
Bombyx mori L. chlorophyll a (VIII) crude extract 100g is dissolved in 500mL ether, adds isopyknic dense salt under the conditions of 0～5 DEG C Reaction 1h is also stirred in acid, divides and takes off a layer acid solution, adds 2 times amount water dilutions, and being neutralized to pH with the NaOH of 10mol/L under cooling is 5-6, Sucking filtration, P2O5Obtain black powder VII crude product 15g after drying, do not do purification and be directly used in next step reaction.
Embodiment 2: the preparation of purpurin 18 (VI)
VII crude product 15g, is dissolved in 50mL oxolane, adds the dilution of 300mL ether, adds the hydroxide of 25% (w/v) Potassium normal propyl alcohol solution 20mL, logical O under the conditions of 0～5 DEG C2Reaction 2h, with water extraction (300mL × 2), 10%H2SO4Neutralize, mistake Filter, P2O5Being dried, silica gel H column chromatography for separation obtains black powder VI 2.7g.
Embodiment 3: chlorin p6The preparation of trimethyl (V)
Compound VI (2.0g, 3.55mmol), is dissolved in 50mL oxolane, adds 200mL methanol, then adds 0.5mol L- 1 sodium hydrate aqueous solution 250mL, is stirred at room temperature and reacts to 698nm absworption peak disappearance, and add water 500ml, is neutralized to dilute hydrochloric acid PH 5～6, ether extraction (3 × 200mL), anhydrous sodium sulfate is dried, and filters, and ether liquid is in the cryosel bath new preparation of upper addition 2.8% (w/v) CH2N2Ether solution 20mL, reacts 5min, washing, recycling design, obtains through silica gel H pillar layer separation after being dried 1.85g black powder V, productivity 83.6%, purity (HPLC): 99.8%.
Embodiment 4: the preparation of chlorin p6 dimethyl ester (IV)
Compound V (375mg, 0.601mmol) is dissolved in 100mL oxolane, adds 25% hydrochloric acid of 100mL, and room temperature is anti- Answer 4 hours.After reaction completely, adding 2.5 times amount water, regulate pH to 5-6 with sodium hydrate aqueous solution, dichloromethane extracts.Have Machine layer is washed 3 times, and anhydrous sodium sulfate is dried, and reduces pressure and steams dichloromethane, silica gel H column chromatography purification (eluant: dichloromethane/ Methanol=100/3), obtain black solid IV (300mg, 0.492mmol), productivity 81.8%.MS(ESI+)m/z:611.39[M+H]+ (100%).UV-visλmax(CH3OH,nm)(ε/M-1cm-1):667(1.73×105),610(2.23×104),529(2.31× 104),497(6.22×104),397(3.47×105)。1H NMR[CDCl3,600MHz]:δ9.69(s,1H),9.47(s,1H), 8.66 (s, 1H), 7.95 (dd, J=18.0,11.4Hz, 1H), 6.29 (d, J=18.0Hz, 1H), 6.12 (d, J=11.4Hz, 1H),5.16(m,1H),4.39(m,1H),4.20(s,3H),4.14(s,3H),3.71(m,2H),3.64(s,3H),3.38(s, 3H), 3.23 (s, 3H), 2.43-2.10 (m, 4H), 1.85 (d, J=6.0Hz, 3H), 1.69 (t, J=7.8Hz, 3H) ,-1.0 (br s,2H).HPLC purity:98.8%.
Embodiment 5:21-bromoethyl-2-goes the preparation of vinyl chlorin p6 dimethyl ester (III)
Compounds Ⅳ (1.0g, 1.64mmol), adds 33%HBr glacial acetic acid liquid 100mL, and room temperature lucifuge seals reaction 36h, subtracts Pressure is evaporated off glacial acetic acid, obtains dark green solid compound III, does not do purification and is directly used in next step reaction.
Embodiment 6:2-(1-methoxyl group) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II1) preparation
Above-mentioned dark green solid compound III (0.1g), is dissolved in dry methylene chloride 50mL, adds K2CO3(100mg) and 2mL is dried methanol, at N2React complete after the lower room temperature reaction 2.5h of protection, stopped reaction, filters, washes 3 times, saturated aqueous common salt Wash 1 time, anhydrous Na2SO4It is dried 2h, silica gel H chromatographic isolation (eluant: methylene chloride/methanol/the third after recovered under reduced pressure organic solvent Ketone/formic acid=100/2/2/0.1, v/v/v/v), obtain black powder II130.4mg, productivity 32.7%.
MS(ESI+)m/z:643.73[M+H]+(100%)；HRMS(ESI+)m/z:642.3059[M]+,calcd for C36H42N4O7 642.3053。
1H NMR[(CD3)2SO,300MHz]:δ9.79(s,2H),8.98(s,1H),5.97(m,1H),4.99(m,1H), 4.55(m,1H),4.13(s,3H),4.10(s,3H),3.73(m,2H),3.60(s,3H),3.47(s,3H),3.30(s,3H), 2.35(m,2H),2.15(m,2H),2.00(d,3H),1.78(d,3H),1.61(m,3H),-1.16(s,1H),-1.30(s, 1H)。
Embodiment 7:2-(1-ethyoxyl) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II2) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried dehydrated alcohol react prepare black solid II2 26.8mg, yield 28.2%.
MS(ESI+)m/z:657.50[M+H]+(100%).
1H NMR[(CD3)2SO,300MHz]:δ9.62(s,1H),9.06(s,1H),8.82(s,1H),8.77(s,1H), 5.91(m,1H),4.92(m,1H),4.51(m,1H),4.32(s,3H),4.15(s,3H),3.69(m,2H),3.58(s,3H), 3.45(s,3H),3.33(s,3H),2.32(m,2H),2.21(m,2H),1.96(d,3H),1.83(d,3H),1.63(m,3H), 0.91(m,3H),-1.01(s,1H),-1.26(s,1H)。
Embodiment 8:2-(1-positive propoxy) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II3) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried anhydrous normal propyl alcohol react prepare black solid II3 31.1mg, yield 32.1%.
MS(ESI+)m/z:671.46[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ12.06(s,1H),9.85(s,1H),9.72(s,1H),8.97(s,1H), 5.96(m,1H),4.99(m,1H),4.57(m,1H),4.14(s,3H),4.12(s,3H),3.69(m,3H),3.57(s,3H), 3.38(s,3H),3.16(s,3H),2.37(m,1H),2.14(m,1H),2.09(m,2H),1.97(d,3H),1.80(d,3H), 1.64(m,3H),1.56(m,3H),0.89(m,3H),-1.13(s,1H),-1.28(s,1H)。
Embodiment 9:2-(1-n-butoxy) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II4) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried anhydrous normal butyl alcohol react prepare black solid II4 27.5mg, yield 27.8%.
MS(ESI+)m/z:685.49[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ12.01(s,1H),9.83(s,1H),9.65(s,1H),8.98(s,1H), 5.88(m,1H),5.05(m,1H),4.58(m,1H),4.16(s,3H),4.15(s,3H),3.58(m,3H),3.55(s,3H), 3.47(m,3H),3.35(s,3H),3.10(d,3H),2.40(m,1H),2.12(m,4H),1.92(d,3H),1.83(d,3H), 1.68(m,3H),0.72(m,3H),-1.13(s,1H),-1.25(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ174.91,174.38,170.22,167.59,166.58,145.46, 141.12,140.85,137.70,136.20,135.17,134.73,132.66,129.09,112.95,104.73,103.70, 101.03,94.46,72.41,68.82,53.24,52.90,52.73,48.81,41.72,34.11,32.22,29.23, 24.83,23.89,19.49,19.09,17.90,14.13,12.49,11.09。
Embodiment 10:2-(1-n-pentyloxy) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II5) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried anhydrous n-amyl alcohol react prepare black solid II5 27.0mg, yield 26.7%.
MS(ESI+)m/z:699.52[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ12.05(s,1H),9.87(s,1H),9.78(s,1H),8.96(s,1H), 5.95(m,1H),4.99(m,1H),4.55(m,1H),4.15(s,3H),4.11(s,3H),3.66(m,3H),3.60(s,3H), 3.49(m,1H),3.38(s,3H),3.19(d,3H),2.36(m,1H),2.06(m,2H),1.99(m,4H),1.80(d,3H), 1.66(m,2H),1.60(m,3H),1.38(m,2H),1.28(m,2H),0.74(m,3H),-1.14(s,1H),-1.27(s, 1H)。
13C NMR[(CD3)2SO,600MHz]:δ174.91,174.38,170.22,167.59,166.58,145.46, 141.12,140.85,137.70,136.20,135.17,134.73,132.66,129.09,112.95,104.73,103.70, 101.03,94.46,72.41,68.82,53.24,52.90,52.73,48.81,41.72,34.11,32.22,29.23, 24.83,23.89,19.49,19.09,17.90,14.13,12.49,11.09。
Embodiment 11:2-(the positive hexyloxy of 1-) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II6) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried anhydrous n-amyl alcohol react prepare black solid II6 37.4mg, yield 36.3%.
MS(ESI+)m/z:713.45[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ12.07(s,1H),9.81(s,1H),9.67(s,1H),8.95(s,1H), 5.87(m,1H),5.02(m,1H),4.56(m,1H),4.15(s,3H),4.13(s,3H),3.55(s,3H),3.51(m,3H), 3.37(m,3H),3.34(s,3H),3.11(d,3H),2.40(m,1H),2.10(m,2H),1.93(d,3H),1.81(d,3H), 1.68(m,3H),1.52(m,3H),0.56(m,3H),-1.13(s,1H),-1.27(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ173.84,173.48,169.72,166.87,166.10,154.25, 148.25,145.10,140.48,137.06,135.85,134.65,134.00,132.10,128.50,122.41,104.30, 103.11,100.67,93.80,71.90,68.61,53.24,52.70,52.38,48.28,31.24,30.95,29.54, 28.76,25.44,24.41,23.36,21.85,18.61,17.45,13.57,11.97,10.57。
Embodiment 12:2-(1-n-octyloxy) ethyl-2-removes vinyl chlorin p6 dimethyl ester (II7) preparation
As described in Example 6, compound III (0.1g) and 2mL be dried anhydrous n-octyl alcohol react prepare black solid II7 38.0mg, yield 35.5%.
MS(ESI+)m/z:741.52[M+H]+(100%).
Embodiment 13:N-(chlorin p6 dimethyl ester-73-acyl group)-Pidolidone (I1) preparation
Compounds Ⅳ (110mg, 0.180mmol) is dissolved in 30mL dry methylene chloride, add EDCI (41.5mg, 0.216mmol, 1.2equiv.) and HOBt (29.2mg, 0.216mmol, 1.2equiv.), the lower stirring of nitrogen protection cryosel bath is anti- Answer 30min, by L-Glu (OBut)2HCl (60.9mg, 0.216mmol, 1.2equiv.) and DIPEA (0.038mL, 0.216mmol, 1.2equiv.) be dissolved in 10mL dichloromethane after, be added dropwise to reactant liquor, under nitrogen protection room temperature condition, stirring is anti- Should be overnight.It is subsequently adding 200mL dichloromethane, washs with 5% aqueous citric acid solution, saturated aqueous common salt and water successively, anhydrous sulfur Acid sodium is dried, recycling design.Solid residue is directly dissolved in 15mL dry methylene chloride, is subsequently adding 5mL trifluoroacetic acid, room temperature Stirring reaction 6 hours.After reaction completely, add 100mL dichloromethane, 10% sodium bicarbonate aqueous solution regulation pH to 3-4, silica gel H column chromatography (eluant: methylene chloride/methanol/acetone/formic acid=100/5/5/0.1), obtains black solid product I1, 82.5mg, 0.112mmol, two step yields 61.9%；HPLC purity:97.6%.
MS(ESI+)m/z:740.42[M+H]+(100%)；HRMS(ESI+)m/z:739.3290[M]+,calcd for C40H45N5O9 739.3217。
UV-visλmax(CH3OH,nm)(ε/M-1cm-1):666(2.72×105),528(5.39×104),497(8.92× 104),397(4.81×105)。
1H NMR[(CD3)2SO,600MHz]:δ12.33(br s,2H),9.67(s,1H),9.45(s,1H),8.98(s, 1H), 8.10 (dd, J=18,12Hz, 1H), 8.03 (d, J=12Hz, 1H), 6.34 (d, J=18Hz, 1H), 6.10 (d, J= 12Hz,1H),4.99(m,1H),4.58(m,1H),4.15(s,3H),4.12(s,3H),3.98(m,2H),3.56(s,3H), 3.40(s,3H),3.38(m,1H),3.09(s,3H),2.28(m,2H),2.18(m,2H),2.01(m,2H),1.88(m,2H), 1.83 (d, J=6.0Hz, 3H), 1.65 (t, J=6.0Hz, 3H) ,-1.14 (s, 1H) ,-1.29 (s, 1H).13C NMR[(CD3)2SO,600MHz]:δ173.59,173.42,173.30,171.74,169.75,167.27,166.09,154.16,148.17, 144.95,140.56,137.28,136.03,135.11,134.20,131.19,128.76,128.66,122.63,122.53, 104.22,103.13,99.79,94.13,52.79,52.62,52.29,51.10,48.16,32.43,31.53,30.01, 28.89,26.22,23.49,18.54,17.51,12.00,11.85,10.64。
Compound I1Chemical structural formula as follows:
[2-(1-methoxyl group) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 14:N-3-acyl group]-L- Glutamic acid (I2) preparation
As described in Example 13, compound ii1With L-Glu (OBut)2HCl prepares black solid I2, yield 61.2%.
MS(ESI+)m/z:772.58[M+H]+(100%).
1H NMR[(CD3)2SO,300MHz]:δ9.81(s,1H),9.79(s,1H),8.96(s,1H),5.97(m,1H), 4.96(m,1H),4.59(m,1H),4.14(s,3H),4.10(s,3H),3.98(m,2H),3.75(m,3H),3.61(s,3H), 3.49(s,3H),3.42(s,3H),3.23(s,3H),2.27-2.11(m,5H),2.00(m,4H),1.81(d,3H),1.63 (m,3H),-1.12(s,1H),-1.29(s,1H)。
Compound I2Chemical structural formula as follows:
[2-(1-ethyoxyl) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 15:N-3-acyl group]-L- Glutamic acid (I3) preparation
As described in Example 13, compound ii2With L-Glu (OBut)2HCl prepares black solid I3, yield 65.3%.
MS(ESI+)m/z:786.56[M+H]+(100%).
Compound I3Chemical structural formula as follows:
[2-(1-positive propoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 16:N-3-acyl group]- Pidolidone (I4) preparation
As described in Example 13, compound ii3With L-Glu (OBut)2HCl prepares black solid I4, yield 63.6%.
MS(ESI+)m/z:800.57[M+H]+(100%)；HRMS(ESI+)m/z:799.3808[M]+,calcd for C43H53N5O10 799.3792。
1H NMR[(CD3)2SO,300MHz]:δ9.86(s,1H),9.78(s,1H),8.95(s,1H),7.71(s,1H), 5.99(m,1H),4.98(m,1H),4.57(m,1H),4.14(s,3H),4.11(s,3H),3.72(m,3H),3.66(m,1H), 3.60(s,3H),3.47(m,2H),3.40(s,3H),3.20(s,3H),2.27-2.03(m,5H),2.00(m,4H),1.81 (d,3H),1.68(m,4H),1.61(m,3H),0.91(m,3H),-1.12(s,1H),-1.27(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ174.84,174.33,171.45,170.21,167.83,166.59, 154.74,148.71,145.73,141.05,140.84,137.62,136.46,135.21,135.11,134.51,132.73, 128.95,122.88,104.88,103.60,101.08,94.33,72.41,70.86,53.26,53.13,52.75,52.42, 48.70,33.18,32.10,31.71,29.41,25.01,23.94,23.40,22.51,19.22,18.11,14.37, 12.54,11.25。
Compound I4Chemical structural formula as follows:
[2-(1-n-butoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 17:N-3-acyl group]- Pidolidone (I5) preparation
As described in Example 13, compound ii4With L-Glu (OBut)2HCl prepares black solid I5, yield 45.0%.
MS(ESI+)m/z:814.53[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ9.90(s,1H),9.81(s,1H),8.99(s,1H),7.96(m,1H), 6.00(m,1H),5.04(m,1H),4.62(m,1H),4.19(s,3H),4.16(s,3H),3.74(m,3H),3.64(s,3H), 3.56(m,1H),3.44(s,3H),3.24(s,3H),2.35-2.12(m,5H),2.03(m,4H),1.86(d,3H),1.70 (m,4H),1.65(m,3H),-1.07(s,1H),-1.22(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ173.76,171.42,169.72,167.25,166.08,154.23, 148.21,145.21,140.53,137.11,135.93,134.59,134.02,132.11,128.47,122.39,104.35, 103.12,100.60,93.79,71.95,68.38,52.74,52.24,51.37,48.21,32.46,31.77,31.53, 31.19,30.52,28.89,26.71,24.45,23.44,19.03,18.71,17.59,13.72,12.03,10.65。
Compound I5Chemical structural formula as follows:
[2-(1-n-pentyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 18:N-3-acyl group]- Pidolidone (I6) preparation
As described in Example 13, compound ii5With L-Glu (OBut)2HCl prepares black solid I6, yield 50.0%.
MS(ESI+)m/z:828.58[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ9.84(s,1H),9.74(s,1H),8.93(s,1H),7.73(m,1H), 5.91(m,1H),4.98(m,1H),4.58(m,1H),4.13(s,3H),4.11(s,3H),3.64(m,4H),3.58(s,3H), 3.46(m,1H),3.37(s,3H),3.17(s,3H),2.28-2.14(m,4H),2.09(m,2H),1.97(d,3H),1.81 (d,3H),1.72(m,2H),1.65(m,4H),1.59(m,3H),0.73(t,3H),-1.12(s,1H),-1.27(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ173.79,171.02,169.72,167.31,166.08,154.20, 148.19,145.18,140.48,137.09,135.88,134.67,134.03,132.18,128.46,122.38,104.32, 103.12,100.62,93.80,71.92,68.65,52.74,52.22,51.90,48.23,32.72,31.62,31.36, 31.16,29.32,28.85,27.57,24.48,23.44,18.68,17.54,13.72,12.01,10.62。
Compound I6Chemical structural formula as follows:
[2-(the positive hexyloxy of 1-) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 19:N-3-acyl group]- Pidolidone (I7) preparation
As described in Example 13, compound ii6With L-Glu (OBut)2HCl prepares black solid I7, yield 48.7%.
MS(ESI+)m/z:842.60[M+H]+(100%).
Compound I7Chemical structural formula as follows:
[2-(1-n-octyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 20:N-3-acyl group]- Pidolidone (I8) preparation
As described in Example 13, compound ii7With L-Glu (OBut)2HCl prepares black solid I8, yield 49.3%.
MS(ESI+)m/z:870.62[M+H]+(100%).
Compound I8Chemical structural formula as follows:
Embodiment 21:N-(chlorin p6 dimethyl ester-73-acyl group)-L-Aspartic acid (I9) preparation
As described in Example 13, compounds Ⅳ and L-Asp (OBut)2HCl prepares I9, yield 58.0%；HPLC Purity:96.3%.
MS(ESI+)m/z:726.43[M+H]+(100%).
UV-visλmax(CH3OH,nm)(ε/M-1cm-1):667(3.73×105),612(5.46×104),529(6.14× 104),497(1.01×105),398(6.15×105)。
1H NMR[(CD3)2SO, 600MHz]: δ 9.48 (s, 1H), 9.19 (s, 1H), 8.95 (s, 1H), 8.05 (d, J= 7.2Hz, 1H), 7.89 (dd, J=18,12Hz, 1H), 6.17 (d, J=18Hz, 1H), 5.96 (d, J=12Hz, 1H), 5.01 (m,1H),4.61(m,1H),4.45(m,2H),4.17(s,3H),4.16(s,3H),3.51(s,3H),3.36(m,1H),3.31 (s, 3H), 2.88 (s, 3H), 2.61 (m, 1H), 2.49 (s, 2H), 2.35 (m, 1H), 2.09 (m, 2H), 1.84 (d, J= 6.6Hz, 3H), 1.44 (t, J=7.2Hz, 3H) ,-1.25 (s, 1H) ,-1.33 (s, 1H).
13C NMR[(CD3)2SO,600MHz]:δ173.37,172.84,172.03,171.24,169.79,167.31, 166.13,154.03,148.05,144.73,140.50,137.17,135.81,134.95,134.21,131.05,128.62, 122.48,122.39,104.05,103.13,99.61,94.10,52.82,52.69,52.28,48.62,48.18,37.31, 32.73,31.64,28.83,23.51,18.41,17.42,11.97,11.77,10.47。HRMS(ESI+)m/z:725.3061 [M]+,calcd for C39H43N5O9 725.3086。
Compound I9Chemical structural formula as follows:
[2-(1-methoxyl group) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 22:N-3-acyl group]-L- Aspartic acid (I10) preparation
As described in Example 13, compound ii1With L-Asp (OBut)2HCl prepares I10, yield 62.2%.
MS(ESI+)m/z:758.51[M+H]+(100%).
1H NMR[(CD3)2SO,300MHz]:δ9.81(s,2H),8.97(s,1H),7.67(m,1H),5.98(m,1H), 4.99(m,1H),4.59(m,1H),4.15(s,3H),4.11(s,3H),3.76(m,3H),3.67(m,1H),3.61(s,3H), 3.48(s,3H),3.42(s,3H),3.23(s,3H),2.34-2.17(m,5H),2.02(m,4H),1.82(d,3H),1.63 (m,3H),-1.12(s,1H),-1.29(s,1H)。
Compound I10Chemical structural formula as follows:
[2-(1-ethyoxyl) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 23:N-3-acyl group]-L- Aspartic acid (I11) preparation
As described in Example 13, compound ii2With L-Asp (OBut)2HCl prepares I11, yield 65.2%.
MS(ESI+)m/z:772.48[M+H]+(100%).
Compound I11Chemical structural formula as follows:
[2-(1-positive propoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 24:N-3-acyl group]- L-Aspartic acid (I12) preparation
As described in Example 13, compound ii3With L-Asp (OBut)2HCl prepares I12, yield 67.0%.
MS(ESI+)m/z:786.44[M+H]+(100%).
1H NMR[(CD3)2SO,300MHz]:δ9.86(s,1H),9.79(s,1H),8.94(s,1H),7.64(m,1H), 6.00(m,1H),4.96(m,1H),4.58(m,1H),4.14(s,3H),4.10(s,3H),3.74(m,3H),3.67(m,2H), 3.60(s,3H),3.49(m,2H),3.40(s,3H),3.21(s,3H),2.38-2.15(m,5H),2.00(m,4H),1.81 (d,3H),1.68(m,2H),1.61(m,3H),0.91(m,3H),-1.13(s,1H),-1.28(s,1H)。
Compound I12Chemical structural formula as follows:
[2-(1-n-butoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 25:N-3-acyl group]- L-Aspartic acid (I13) preparation
As described in Example 13, compound ii4With L-Asp (OBut)2HCl prepares I13, yield 77.1%.
MS(ESI+)m/z:800.68[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ9.90(s,1H),9.80(s,1H),8.99(s,1H),7.75(m,1H), 6.00(m,1H),5.03(m,1H),4.64(m,1H),4.28,(m,1H),4.19(s,3H),4.17(s,3H),3.74(m, 2H),3.64(s,3H),3.56(m,2H),3.44(s,3H),3.23(s,3H),2.50-2.30(m,4H),2.10(m,2H), 2.03(m,5H),1.86(d,3H),1.70(m,2H),1.65(m,6H),-1.07(s,1H),-1.21(s,1H)。
13C NMR[(CD3)2SO,600MHz]:δ173.81,170.62,169.69,167.33,166.09,154.22, 148.20,145.18,140.50,137.08,135.90,134.59,134.03,132.17,128.46,122.37,104.34, 103.10,100.58,93.83,71.95,68.37,52.77,52.24,48.60,48.22,32.78,31.76,31.64, 31.17,28.86,28.57,24.49,23.43,21.99,19.03,18.70,17.57,13.71,12.03,10.77。
Compound I13Chemical structural formula as follows:
[2-(1-n-pentyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 26:N-3-acyl group]- L-Aspartic acid (I14) preparation
As described in Example 13, compound ii5With L-Asp (OBut)2HCl prepares I14, yield 50.2%.
MS(ESI+)m/z:814.47[M+H]+(100%).
1H NMR[(CD3)2SO,600MHz]:δ9.83(s,1H),9.69(s,1H),8.96(s,1H),7.95(m,1H), 5.91(m,1H),5.00(m,1H),4.60(m,1H),4.37(s,1H),4.15(s,3H),4.14(s,3H),3.59(m,4H), 3.56(s,3H),3.43(m,2H),3.37(s,3H),3.14(s,3H),2.57(m,1H),2.09(m,2H),1.96(d,3H), 1.83(d,3H),1.72(m,2H),1.62(m,4H),1.58(m,3H),0.70(t,3H),-1.10(s,1H),-1.25(s, 1H)。
13C NMR[(CD3)2SO,600MHz]:δ173.78,172.98,172.19,171.03,169.73,167.28, 166.10,154.21,148.19,145.14,140.54,140.32,137.07,135.85,134.67,134.04,132.15, 128.47,122.40,104.27,103.14,100.63,93.81,71.93,68.63,52.77,52.22,48.60,48.24, 32.67,31.60,29.30,28.78,28.04,24.45,23.43,21.86,18.64,17.50,13.69,11.99, 10.74。
Compound I14Chemical structural formula as follows:
[2-(the positive hexyloxy of 1-) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 27:N-3-acyl group]- L-Aspartic acid (I15) preparation
As described in Example 13, compound ii6With L-Asp (OBut)2HCl prepares I15, yield 48.4%.
MS(ESI+)m/z:828.57[M+H]+(100%).
Compound I15Chemical structural formula as follows:
[2-(1-n-octyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 28:N-3-acyl group]- L-Aspartic acid (I16) preparation
As described in Example 13, compound ii7With L-Asp (OBut)2HCl prepares I16, yield 46.8%.
MS(ESI+)m/z:856.61[M+H]+(100%).
Compound I16Chemical structural formula as follows:
Embodiment 29:N-chlorin p6 dimethyl ester-73-acyl group)-1B (I17) preparation
Compounds Ⅳ (110mg, 0.180mmol) is dissolved in 30mL dry methylene chloride, add EDCI (41.5mg, 0.216mmol, 1.2equiv.) and HOBt (29.2mg, 0.216mmol, 1.2equiv.), the lower stirring of nitrogen protection cryosel bath is anti- Answer 30min, by Nε-tertbutyloxycarbonyl-1B methyl ester hydrochloride (Nε-Boc-L-Lys-OMe·HCl)(64.1mg, 0.216mmol, 1.2equiv.) and after DIPEA (0.038mL, 0.216mmol, 1.2equiv.) is dissolved in 10mL dichloromethane, drip Adding reactant liquor, under nitrogen protection room temperature condition, stirring reaction is overnight.It is subsequently adding 200mL dichloromethane, uses 5% Fructus Citri Limoniae successively Aqueous acid, saturated aqueous common salt and water washing, anhydrous sodium sulfate is dried, recycling design.Solid residue is dissolved in 15mL THF, adds LiOH (33.6mg, 1.4mmol, 8.0equiv.) aqueous solution (5mL) and methanol (10mL), be stirred at room temperature reaction 2h, reacted Finish.Reactant liquor add water (50mL) dilution, use saturated NH4Cl liquid regulation pH to 5-6, extracts (20mL × 3) with dichloromethane, anhydrous Na2SO4Being dried, decompression removes dichloromethane, P2O5It is vacuum dried to obtain black solid.Above-mentioned black solid is redissolved in 15mL and does Dry dichloromethane, adds 5mL trifluoroacetic acid (TFA) reaction 1h, adds 85mL dchloromethane after completion of the reaction, with saturated under ice bath NaHCO3Neutralizing unreacted TFA is puce to reactant liquor by green transition, point takes organic layer, recovered under reduced pressure organic solvent, warp Silica gel H pillar layer separation, gradient elution agent CH2Cl2: MeOH=4:1～2:1 (v/v), obtain black powder I1756,5mg, yield 42.50%.
MS(ESI+)m/z:739.43[M+H]+(100%).
Compound I17Chemical structural formula as follows:
[2-(1-methoxyl group) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 30:N-3-acyl group]-L- Lysine (I18) preparation
By the method for embodiment 29, compound ii1With Nε-Boc-L-Lys-OMe HCl prepares I18, yield 45.2%.
MS(ESI+)m/z:771.48[M+H]+(100%).
Compound I18Chemical structural formula as follows:
[2-(1-ethyoxyl) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 31:N-3-acyl group]-L- Lysine (I19) preparation
By the method for embodiment 29, compound ii2With Nε-Boc-L-Lys-OMe HCl prepares I19, yield 40.8%.
MS(ESI+)m/z:785.51[M+H]+(100%).
Compound I19Chemical structural formula as follows:
[2-(1-positive propoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 32:N-3-acyl group]- 1B (I20) preparation
By the method for embodiment 29, compound ii3With Nε-Boc-L-Lys-OMe HCl prepares I20, yield 47.4%.
MS(ESI+)m/z:799.54[M+H]+(100%).
Compound I20Chemical structural formula as follows:
[2-(1-n-butoxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 33:N-3-acyl group]- 1B (I21) preparation
By the method for embodiment 29, compound ii4With Nε-Boc-L-Lys-OMe HCl prepares I21, yield 48.5%.
MS(ESI+)m/z:813.43[M+H]+(100%).
Compound I21Chemical structural formula as follows:
[2-(1-n-pentyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 34:N-3-acyl group]- 1B (I22) preparation
By the method for embodiment 29, compound ii5With Nε-Boc-L-Lys-OMe HCl prepares I22, yield 38.2%.
MS(ESI+)m/z:827.58[M+H]+(100%).
Compound I22Chemical structural formula as follows:
[2-(the positive hexyloxy of 1-) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 35:N-3-acyl group]- 1B (I23) preparation
By the method for embodiment 29, compound ii6With Nε-Boc-L-Lys-OMe HCl prepares I23, yield 43.7%.
MS(ESI+)m/z:841.47[M+H]+(100%).
Compound I23Chemical structural formula as follows:
[2-(1-n-octyloxy) ethyl-2-removes vinyl-chlorin p6 dimethyl ester-7 to embodiment 36:N-3-acyl group]- 1B (I24) preparation
By the method for embodiment 29, compound ii7With Nε-Boc-L-Lys-OMe HCl prepares I24, yield 47.6%.
MS(ESI+)m/z:869.63[M+H]+(100%).
Compound I24Chemical structural formula as follows:
Embodiment 37: the external PDT anti-tumor activity examination of part of the present invention preferred chlorin p6 amino acid derivant Test.
1. material
Cell strain selects mouse melanin tumor cell (B16-F10) and human melanoma cell (A375), purchased from section of China Institute's Shanghai cell bank.
LASER Light Source uses the 660nm semiconductor laser therapeutic instrument that Ennai Laser Technology Co., Ltd., Shanghai develops, Big output is 2W.
2. method
1) cell is cultivated: B16-F10 and A375 cell is incubated at respectively and adds the sodium bicarbonate of 2mg/mL, 4.5mg/mL Glucose, 100 μ g/mL streptomycin sulfates, 40mg/mL gentamycin, 100U/mL penicillin, 10% (v/v) heat inactivation tire cattle The RPMI 1640 culture medium (Hyclone, Logan, UT, USA) of serum (FBS) or DMEM culture medium (Hyclone, Logan, UT, USA), it is placed in the constant incubator that 5% carbon dioxide temperature is 37 DEG C cultivation.
2) drug solution preparing: take testing compound of the present invention and positive control medicine Verteporfin (verteporfin, BPD- MA) being dissolved in DMSO, the medicine being made into 300mM concentration stores liquid.
3) the dark toxicity of cell: (Costar, Cambridge, MA, USA) inoculates 5 × 10 on the 96 every holes of orifice plate3Individual B16- F10 or A375 cell, be then incubated at above-mentioned under the conditions of cultivate.Take testing sample and be made into the liquid to be measured of concentration known, be diluted to Variable concentrations add cell culture fluid, after be further cultured for 48 hours.
CCK-8 algoscopy: by cell culture fluid sucking-off, adds containing 10% (v/v) CCK-8 (Dojindo Laboratories, Japan) culture fluid 200 μ L, continue cultivate 1.5 hours, then with microplate reader (Tecan, Switzerland) wavelength 450nm, measures the absorbance in every hole.
4) cell phototoxicity: B16-F10 or A375 cell is according to above-mentioned CMC model, and add concentration known treats test sample Product, cultivate 24 hours.Then 660nm wavelength light irradiation, light dosage 10J/cm are used2, continue to cultivate 24 hours.Select CCK-8 again Method measures the survival rate of cell.
3. result
Moiety intermediate of the present invention and the dark toxicity of selected objective target Compounds in vitro tumor cell and PDT lethal effect are tied Really, 2 it are shown in Table.
Table 2 moiety intermediate and the target compound half-inhibition concentration IC to tumor cell50(μM)
Above-mentioned experimental result shows:
It is 10J/cm at illumination dose2Under the conditions of, tested tumor cell line is respectively provided with excellent by most test compounds Good PDT active anticancer, and its therapeutic index (dark poison and light poison than) is significantly better than the positive control photosensitive drug of similar listing Verteporfin (verteporfin, BPD-MA).
Below preferred embodiment to the invention is illustrated, but the invention is not limited to described Embodiment, those of ordinary skill in the art also can make all equivalents on the premise of the invention spirit Modification or replacement, modification or the replacement of these equivalents are all contained in the application claim limited range.

Claims (10)

1. chlorin p6 amino acid derivant and a pharmaceutical salts thereof, described chlorin p6 amino acid derivant Chemical constitution is as shown in formula I:
In formula I,
R1Represented amino acid residue；
R2Represent CH=CH2、CH(OR3)CH3；
R3Represent H, low alkyl group, senior alkyl, (CH2)mOR4Or (CH2CH2O)kR5；
R4Represent H, low alkyl group；
R5Represent H, low alkyl group；
M represents the integer between 3-6；
K represents the integer between 1-6；
Described low alkyl group, refers to the straight or branched alkyl containing 1-6 carbon atom；
Described senior alkyl, refers to the straight or branched alkyl containing 7-18 carbon atom.
A kind of chlorin p6 amino acid derivant the most according to claim 1 and pharmaceutical salts thereof, it is characterised in that formula (I) in,
R1Represent lysine, glutamic acid or aspartic acid；
R2Represent CH=CH2、CH(OCH3)CH3、CH(OC2H5)CH3、CH(O-n-C3H7)CH3、CH(O-n-C4H9)CH3、CH(O-n- C5H11)CH3、CH(O-n-C6H11)CH3、CH(O-n-C8H17)CH3。
A kind of chlorin p6 amino acid derivant the most according to claim 2 and pharmaceutical salts thereof, it is characterised in that institute R in the chlorin p6 amino acid derivant stated1And R2It is respectively combined as follows:
4., according to a kind of chlorin p6 amino acid derivant described in claim 1,2 or 3 and pharmaceutical salts thereof, its feature exists In, described pharmaceutical salts is alkali metal salt.
A kind of chlorin p6 amino acid derivant the most as claimed in claim 1 and the preparation method of pharmaceutical salts thereof, it is special Levying and be, the method comprises the following steps:
With purpurin 18 as raw material, the chlorin p6 bis-prepared through the hydrolysis of inner-acid anhydride alkali open ring, carboxymethyl group and ester successively Methyl ester is key intermediate, 7-the third carboxyl in chlorin p6 dimethyl ester structure and amino acid condensation or its 2-vinyl and 7- Third carboxyl respectively with alcohol and aminoacid generation addition and condensation reaction, synthesizing dihydro porphin p6 amino acid derivant.
A kind of chlorin p6 amino acid derivant the most according to claim 5 and the preparation method of pharmaceutical salts thereof, its Being characterised by, described synthesis material purpurin 18 chlorophyll a prepares through acid, alkali and oxidative degradation.
A kind of chlorin p6 amino acid derivant the most according to claim 5 and the preparation method of pharmaceutical salts thereof, its Being characterised by, the method comprises the following steps:
A, it is that chlorin p6 dimethyl ester (chlorin p6 prepared by raw material with chlorophyll a (chlorophyll a, VIII) Dimethylester, IV), reaction process is as follows, and reaction method is A1 to A2:
A1, prepare purpurin 18 (purpurin-18, VI)
First, the diethyl ether solution of commercially available chlorophyll paste, with equal volume amounts concentrated hydrochloric acid stirring reaction 0.5h under the conditions of 0～5 DEG C Generate pheophorbide acid a (pheophorbide a, VII)；Then, compound VII is logical in the normal propyl alcohol liquid of 25% potassium hydroxide Oxygen reaction 2 prepares VI；
A2, prepare chlorin p6 dimethyl ester (chlorin p6 dimethylester, IV)
Compound VI is dissolved in oxolane-methanol (1:4, v/v), adds equal-volume 0.5mol.L-1 sodium hydroxide reaction 2h and prepares not Stable product chlorin p6 (chlorine p6) is rapid afterwards and diazomethane reaction prepares chlorin p6 trimethyl (chlorin p6 trimethylester, V), compound V prepares IV through 25%HCl hydrolysis；
B, preparing target compound chlorin p6 amino acid derivatives I with compounds Ⅳ, reaction process is as follows, reaction method For B1 to B2:
B1, prepare chlorin p6 ether derivative II
The 33%HBr glacial acetic acid liquid room temperature reaction 24h of compounds Ⅳ with excess is prepared intermediate III, and intermediate III is in excess K2CO3In the presence of with various alcohol (R3OH) reaction prepares compound ii；
B2, prepare chlorin p6 amino acid target derivative I:
By compounds Ⅳ or II dichloromethane solution at 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDCI), in the presence of N-hydroxybenzotriazole (HOBt) and N, N-dimethyl isopropyl amine (DIPEA) respectively with various carboxyls or/ Acidity or hydrochloride of basic amino acid (R with amido protecting1'-NH2HCl) slough through trifluoracetic acid (TFA) again after reaction 24h The tert-butyl group or tertbutyloxycarbonyl or and Lithium hydrate demethylation ester prepare chlorin p6 amino acid target derivative I.
8. a kind of chlorin p6 amino acid derivant and pharmaceutical salts thereof as described in claim 1,2 or 3 are anti-swollen in preparation Application in tumor medicine.
A kind of chlorin p6 amino acid derivant the most according to claim 8 and pharmaceutical salts thereof are preparing antineoplastic agent Application in thing, it is characterised in that described antitumor drug is Photodynamic therapy of cancer medicine.
10. chlorin p6 amino acid derivant as claimed in claim 1 a kind of and pharmaceutical salts thereof are controlled preparing light power Treat the application in senile degeneration of macula, nevus flammeus, or condyloma acuminatum medicine.
CN201610356755.4A 2016-05-25 2016-05-25 Chlorin p6 amino acid derivative, preparation method therefor and use of chlorin p6 amino acid derivative Pending CN106046008A (en)

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Publication number Priority date Publication date Assignee Title
CN107987081A (en) * 2016-10-26 2018-05-04 刘辉 A kind of new chlorin e 6 derivative and its pharmaceutically acceptable salt, its preparation method and application
CN107987081B (en) * 2016-10-26 2019-12-06 刘辉 Chlorin e6 derivative and pharmaceutically acceptable salt thereof, and preparation method and application thereof
CN111943954A (en) * 2020-08-18 2020-11-17 广州易斯生物科技发展有限公司 Dihydroporphin derivative and corresponding preparation method and application thereof
CN111943954B (en) * 2020-08-18 2021-08-13 广州易斯生物科技发展有限公司 Dihydroporphin derivative and corresponding preparation method and application thereof
WO2022037599A1 (en) * 2020-08-18 2022-02-24 广州易斯生物科技发展有限公司 Chlorin derivative or pharmaceutically acceptable salt thereof, preparation method therefor, use thereof, and combination thereof with ultrasound medical system
CN112321597A (en) * 2020-11-25 2021-02-05 桂林市兴达光电医疗器械有限公司 Chlorin e6Derivative, preparation method and application thereof
CN113527319A (en) * 2021-06-10 2021-10-22 中国人民解放军海军军医大学 Novel chlorin e4Derivative and pharmaceutically acceptable salt thereof, and preparation method and application thereof
CN113527319B (en) * 2021-06-10 2023-03-07 中国人民解放军海军军医大学 Novel chlorin e4 derivative and pharmaceutically acceptable salt thereof, and preparation method and application thereof

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