CN110498829B - Triptolide derivative, preparation method thereof, pharmaceutical composition thereof and application thereof - Google Patents

Triptolide derivative, preparation method thereof, pharmaceutical composition thereof and application thereof Download PDF

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CN110498829B
CN110498829B CN201810464842.0A CN201810464842A CN110498829B CN 110498829 B CN110498829 B CN 110498829B CN 201810464842 A CN201810464842 A CN 201810464842A CN 110498829 B CN110498829 B CN 110498829B
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triptolide
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张东明
陈晓光
臧应达
来芳芳
李创军
王雨辰
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Abstract

The invention discloses triptolide derivatives, a preparation method thereof, a pharmaceutical composition thereof and application thereof. Specifically, the invention discloses triptolide derivatives shown as a general formula I, II and a compound LA-67. The derivatives are prepared by artificial synthesis, pharmaceutical compositions containing the derivatives and application of the derivatives in preparing medicaments for treating tumor diseases.

Description

Triptolide derivative, preparation method thereof, pharmaceutical composition thereof and application thereof
Technical Field
The invention relates to the field of natural medicines and medicinal chemistry, in particular to triptolide derivatives. The invention also discloses a preparation method of the derivative, a pharmaceutical composition and an application of the derivative in the aspect of tumor resistance.
Background
Tripterygium wilfordii is a traditional Chinese herbal medicine with rich resources owned by China. The triptolide as the main active component is an abietane diterpenoid compound containing 3 epoxy groups and 1 alpha, beta-unsaturated five-membered lactone ring structure, and has obvious biological activities of resisting inflammation, resisting tumor, suppressing immunity, resisting male fertility and the like. Since the discovery, has attracted a great deal of attention from many medicinal chemists and pharmacologists. However, the compound has high toxicity, so that the clinical application of the compound is limited. Therefore, how to improve the safety window of the compounds on the premise of keeping clinical curative effect is a bottleneck of research and development in the field at present.
Nitric oxide has a dual role in anti-tumor, depending mainly on the concentration, time and site of action of nitric oxide production. Sustained low concentrations of nitric oxide promote tumor cell growth, while high concentrations of nitric oxide exert anti-tumor activity by producing cytotoxicity. Furazan nitroxides as nitric oxide donors provide sustained release of high concentrations of nitric oxide in vivo (Kerwin j.f., Heller m., med.res.rev.,1994,14(1): 23.). In addition, the conjugates of nitric oxide donors and drugs often have the effects of synergy and attenuation, and thus are more concerned in the research of new drugs. Therefore, according to the twin drug design principle, the furazan nitrogen oxide and the 14-hydroxyl of triptolide are coupled to obtain the novel anti-tumor drug with an expanded safety window.
Disclosure of Invention
The invention aims to couple the 14-site hydroxyl of triptolide with benzenesulfonyl furazan oxynitride through a connecting bond to obtain a novel anti-tumor drug with an expanded safety window.
Another object of the present invention is to provide a process for producing the above compound.
A further object of the present invention is to provide the use of the above compound or a pharmaceutically acceptable salt thereof.
The object of the invention can be achieved by the following measures:
a compound having the structure of formula I or a pharmaceutically acceptable salt thereof,
Figure BDA0001661798100000011
R1is selected from (CH)2)n(n-1, 2, 3, 4, 5, 6, 7 or 8, C6H4、CH＝CH、CH2C(CH3)2CH2；R2Is selected from (CH)2)n(n-1, 2, 3, 4, 5, 6, 7 or 8), CH (CH)3)CH2CH2、H2CC≡CCH2、(CH2)2O(CH2)2。
The compounds of the invention of formula I can be prepared by the following method:
Figure BDA0001661798100000021
wherein, the condition a: reactant HOR2OH, 30 percent NaOH and tetrahydrofuran as a solvent are reacted at normal temperature; condition b: reacting a reactant which is anhydride, a catalyst which is DMAP and a solvent which is dichloromethane or tetrahydrofuran at normal temperature; condition c: catalysisThe agent is EDCI, HOBt and the solvent is dichloromethane.
In intermediates 1-2a, R1＝CH2CH2，R2＝CH(CH3)CH2CH2；
In intermediates 1-2b, R1＝CH2CH2，R2＝(R)-CH(CH3)CH2CH2；
In intermediates 1-2c, R1＝CH2CH2，R2＝(S)-CH(CH3)CH2CH2；
In intermediates 1-2d, R1＝CH2CH2，R2＝(CH2)4；
In intermediates 1-2e, R1＝CH2CH2，R2＝H2CC≡CCH2；
In intermediates 1-2f, R1＝CH2CH2，R2＝(CH2)2O(CH2)2；
In intermediates 1-2g, R1＝CH2CH2，R2＝(CH2)3；
In intermediates 1-2h, R1＝CH2CH2，R2＝(CH2)5；
In intermediates 1-2i, R1＝(CH2)3，R2＝CH(CH3)CH2CH2；
In intermediates 1-2j, R1＝(CH2)3，R2＝(R)-CH(CH3)CH2CH2；
In intermediates 1-2k, R1＝(CH2)3，R2＝(S)-CH(CH3)CH2CH2；
In intermediates 1-2l, R1＝(CH2)3，R2＝CH2CH2CH2CH2；
In intermediates 1 to 2m, R1＝(CH2)3，R2＝H2CC≡CCH2。
Processes for the preparation of intermediates 1a-m and 2a-m are described in the document J.Med.chem.2008,51, 4834-4838.
A compound having the structure of formula II or a pharmaceutically acceptable salt thereof,
Figure BDA0001661798100000031
R3is selected from (CH)2)n、CH2O(CH2)2、C≡CCH2(ii) a n is 1,2, 3, 4, 5, 6, 7 or 8.
The compounds of the invention of formula II can be prepared by the following process:
Figure BDA0001661798100000032
wherein, the condition a: reactant HOR2OH, 30 percent NaOH and tetrahydrofuran as a solvent are reacted at normal temperature; condition d: the catalyst is Jones reagent (namely chromium trioxide solution in concentrated sulfuric acid), and the reaction is carried out at normal temperature. Condition c: the catalyst is EDCI, HOBt, and the solvent is dichloromethane.
In intermediate 3-4a, R3＝(CH2)3；
In intermediates 3-4b, R3＝(CH2)2OCH2；
In intermediates 3-4c, R3＝(CH2)4；
In intermediates 3 to 4d, R3＝(CH2)5。
The intermediates 3a-d and 4a-d are prepared according to the methods described in the documents J.Med.chem.2015,58, 4325-4338.
A compound having 14-O- (3-benzenesulfonyl-2-oxo-furazan-4-oxo) -triptolide (LA-67) structure or its pharmaceutically acceptable salt,
Figure BDA0001661798100000033
one preparation method of the compound LA-67 is as follows: and reacting triptolide with benzenesulfonyl furazan oxynitride under an alkaline condition to obtain the compound. The reaction equation is as follows:
Figure BDA0001661798100000034
wherein the reaction solvent is tetrahydrofuran or N, N-dimethylformamide; the alkali is sodium hydride or sodium hydroxide; the reaction temperature was 0 ℃.
Some of the compounds of the invention are selected from:
Figure BDA0001661798100000041
in another aspect, the invention relates to pharmaceutical compositions comprising as active ingredient a compound of the invention. The pharmaceutical compositions are prepared according to methods well known in the art. The compounds of the invention may be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The compounds of the present invention are generally present in the pharmaceutical compositions in an amount of 0.1 to 95% by weight.
The compound of the invention or the pharmaceutical composition containing the same can be administrated in a unit dosage form, and the administration route can be intestinal tract or parenteral tract, such as oral administration, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavity, oral mucosa, eyes, lungs, respiratory tract, skin, vagina, rectum and the like.
The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.
The compound can be prepared into common preparations, sustained-release preparations, controlled-release preparations, targeting preparations and various microparticle drug delivery systems.
For tableting the compounds of the invention, a wide variety of excipients well known in the art may be employed, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannose, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like.
The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.
To encapsulate the administration units, the active ingredient of the compounds of the invention can be mixed with diluents and glidants and the mixture can be placed directly into hard or soft capsules. Or the effective component of the compound of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants, glidants used to prepare tablets of the compounds of the present invention may also be used to prepare capsules of the compounds of the present invention.
For preparing the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixture can be used as solvent, and appropriate amount of solubilizer, cosolvent, pH regulator, and osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent may be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc. The pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.
In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.
For the purpose of administration and enhancing the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.
The dosage of the pharmaceutical composition of the compound of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is from 0.001 to 150mg/kg body weight, preferably from 0.1 to 100mg/kg body weight, more preferably from 1 to 60mg/kg body weight, most preferably from 2 to 30mg/kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.
The compounds or compositions of the present invention may be administered alone or in combination with other therapeutic or symptomatic agents. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.
The compound or the pharmaceutically acceptable salt thereof can be applied to the field of pharmacy, and in the field of antitumor drugs, such as in-vivo pharmacodynamic experiments for preparing anti-melanoma drugs, the compound LA-67 oral preparation disclosed by the inventionWhen the dosage is 0.3mg/kg, the compound has remarkable anti-melanoma activity, and the inhibition rate is about 73.5%. Preliminary oral acute toxicity experiments showed that half of the Lethal Dose (LD) of the inventive compound LA-6750) 160.85mg/kg, and LD of triptolide50About 1 mg/kg. The above results indicate that the toxicity of triptolide derivative LA-67 is significantly reduced compared to triptolide, and that it has anti-melanoma activity at low concentrations.
Drawings
FIG. 1 tumor tissue
Detailed Description
EXAMPLE 1 preparation of 14-O- {4- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -4-oxobutanoyl } -triptolide (LA-50)
50mg of triptolide (1equiv) was completely dissolved in 5ml of dichloromethane, and 86mg of intermediate 2a (1.5equiv), 53mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (2equiv, EDCI) and 38mg of 1-hydroxybenzotriazole (2equiv, HOBt) were added to a bottle and stirred at room temperature overnight. Stopping reaction, adding dichloromethane for dilution, washing with a small amount of water and saturated sodium chloride solution once respectively, drying with anhydrous sodium sulfate, filtering, and spin-drying the filtrate to obtain a crude product. Purification was performed by silica gel column chromatography (chloroform: methanol 20:1) to give a white solid, and recrystallization was performed with methanol to give pure 75.6mg, yield 72%. H1NMR(400MHz,CDCl3)δ8.05-8.03(d,J＝7.8Hz,2H),7.75-7.72(t,1H),7.63-7.59(t,2H),5.03(s,1H),4.69-4.60(q,2H),3.81-3.80(d,J＝2.2Hz,1H),3.50-3.49(d,J＝2.6Hz,1H),3.43-3.42(d,J＝5.5Hz,1H),1.32-1.30(d,J＝6.3Hz,3H),1.23(s,2H),1.02(s,3H),0.97-0.95(d,J＝6.8Hz,1H),0.88-0.87(d,J＝6.9Hz,3H),0.79-0.77(d,J＝6.9Hz,3H).HRESIMS m/z＝779.2092[M+Na]+(calcd for C36H40N2NaO14S,779.2092)。
EXAMPLE 2 preparation of (R) -14-O- {4- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -4-oxobutanoyl } -triptolide (LA-51)
Prepared from triptolide and intermediate 2b according to the preparation method of example 2The rate was 58%. H1NMR(400MHz,CDCl3)δ8.05-8.03(d,J＝7.8Hz,2H),7.75-7.72(t,1H),7.63-7.59(t,2H),5.03(s,1H),4.69-4.60(q,2H),3.81-3.80(d,J＝2.2Hz,1H),3.50-3.49(d,J＝2.6Hz,1H),3.43-3.42(d,J＝5.5Hz,1H),1.32-1.30(d,J＝6.3Hz,3H),1.23(s,2H),1.02(s,3H),0.97-0.95(d,J＝6.8Hz,1H),0.88-0.87(d,J＝6.9Hz,3H),0.79-0.77(d,J＝6.9Hz,3H).HRESIMS m/z＝779.2092[M+Na]+(calcd for C36H40N2NaO14S,779.2092)。
EXAMPLE 3 preparation of (S) -14-O- {4- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -4-oxobutanoyl } -triptolide (LA-52)
Prepared from triptolide and intermediate 2c in 63% yield according to the preparation method of example 2. H1NMR(400MHz,CDCl3)δ8.05-8.03(d,J＝7.8Hz,2H),7.75-7.72(t,1H),7.63-7.59(t,2H),5.03(s,1H),4.69-4.60(q,2H),3.81-3.80(d,J＝2.2Hz,1H),3.50-3.49(d,J＝2.6Hz,1H),3.43-3.42(d,J＝5.5Hz,1H),1.32-1.30(d,J＝6.3Hz,3H),1.23(s,2H),1.02(s,3H),0.97-0.95(d,J＝6.8Hz,1H),0.88-0.87(d,J＝6.9Hz,3H),0.79-0.77(d,J＝6.9Hz,3H).HRESIMS m/z＝779.2092[M+Na]+(calcd for C36H40N2NaO14S,779.2092)。
EXAMPLE 4 preparation of 14-O- {4- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -butoxy ] -4-oxobutanoyl } -triptolide (LA-53)
Prepared from triptolide and intermediate 2d in 69% yield according to the preparation method of example 2. H1NMR(400MHz,CDCl3)δ8.07-8.05(d,J＝7.8Hz,2H),7.78-7.75(t,1H),7.65-7.61(t,2H),5.09(s,1H),4.71-4.62(q,2H),4.47-4.44(t,2H),4.22-4.17(m,2H),3.83-3.82(d,J＝3.0Hz,1H),3.53(d,J＝2.3Hz,1H),3.46-3.45(d,J＝5.6Hz,1H),1.05(s,3H),0.96-0.95(d,J＝6.9Hz,3H),0.85-0.83(d,J＝6.9Hz,3H).HRESIMS m/z＝779.2092[M+Na]+(calcd for C36H40N2NaO14S,779.2092)
EXAMPLE 5 preparation of 14-O- {4- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -2-yne-butoxy ] -4-oxobutanoyl } -triptolide (LA-54)
Prepared from triptolide and intermediate 2e in 73% yield according to the preparation method of example 2. H1NMR(400MHz,CDCl3)δ8.07-8.05(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.65-7.61(t,2H),5.09(s,2H),5.07(s,1H),4.76(s,2H),4.66(s,2H),3.82-3.81(d,J＝3.0Hz,1H),3.53-3.52(d,J＝2.3Hz,1H),3.45-3.44(d,J＝5.6Hz,1H),1.58-1.54(m,1H),1.24(s,2H),1.04(s,3H),0.95-0.93(d,J＝6.9Hz,3H),0.84-0.82(d,J＝6.9Hz,3H).HRESIMS m/z＝753.1977[M+H]+(calcd for C36H36N2O14S,752.1887).
EXAMPLE 6 preparation of 14-O- {4- {2- {2- [ 2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) ethoxy ] } -ethoxy } -4-oxobutanoyl } -triptolide (LA-55)
According to the preparation method of example 2, it is prepared from triptolide and intermediate 2f in 75% yield. H1NMR(400MHz,CDCl3)δ8.07-8.05(d,J＝7.8Hz,2H),7.78-7.75(t,1H),7.65-7.61(t,2H),5.07(s,1H),4.71-4.62(q,2H),4.57-4.55(t,2H),3.83-3.82(d,J＝3.0Hz,1H),3.52(d,J＝2.3Hz,1H),3.46-3.45(d,J＝5.6Hz,1H),1.04(s,3H),0.95-0.94(d,J＝6.9Hz,3H),0.84-0.82(d,J＝6.9Hz,3H).HRESIMS m/z＝796.2076[M+Na]+(calcd for C36H40N2O15S,772.2149).
EXAMPLE 7 preparation of 14-O- {4- [3- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -propoxy ] -4-oxobutanoyl } -triptolide (LA-56)
According to the preparation method of example 2, the extract was prepared from triptolide and intermediate 2g in 74% yield. H1NMR(400MHz,CDCl3)δ8.05-8.03(d,J＝7.8Hz,2H),7.77-7.73(t,1H),7.64-7.60(t,2H),5.05(s,1H),4.70-4.60(q,2H),4.54-4.48(t,2H),3.82-3.83(d,J＝3.0Hz,1H),3.52-3.51(d,J＝2.3Hz,1H),3.44-3.43(d,J＝5.6Hz,1H),1.73(s,1H),1.56-1.52(m,1H),1.03(s,3H),0.98-0.96(d,J＝6.7Hz,1H),0.94-0.92(d,J＝6.9Hz,3H),0.82-0.81(d,J＝6.9Hz,3H).HRESIMS m/z＝765.1934[M+Na]+(calcd for C35H38N2O14S,742.2044).
EXAMPLE 8 preparation of 14-O- {4- [5- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -pentyloxy ] -4-oxobutanoyl } -triptolide (LA-57)
According to the preparation method of example 2, the preparation method is prepared from triptolide and an intermediate for 2h, and the yield is 58%. H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.64-7.60(t,2H),5.08(s,1H),4.70-4.60(q,2H),4.44-4.41(t,2H),4.16-4.07(m,4H),3.82-3.81(d,J＝3.0Hz,1H),3.52(d,J＝2.3Hz,1H),3.45-3.44(d,J＝5.6Hz,1H),1.04(s,3H),0.99-0.97(d,J＝6.7Hz,1H),0.96-0.94(d,J＝6.9Hz,3H),0.94-0.92(d,J＝6.9Hz,3H),0.84-0.82(d,J＝6.9Hz,3H).HRESIMS m/z＝793.2247[M+Na]+(calcd for C37H42N2O14S,793.2249).
EXAMPLE 9 preparation of 14-O- {5- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -5-oxovaleryl } -triptolide (LA-58)
Prepared from triptolide and intermediate 2i in 63% yield according to the preparation method of example 2. H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.64-7.60(t,2H),5.06(s,1H),4.66(s,3H),4.46-4.43(m,2H),3.81-3.80(d,J＝3.0Hz,1H),3.51-3.50(d,J＝2.3Hz,1H),3.45-3.44(d,J＝5.6Hz,1H),1.31-1.30(d,J＝6.4Hz,3H),0.93-0.92(d,J＝6.9Hz,3H),0.87-0.85(d,J＝6.9Hz,1H),0.82-0.80(d,J＝6.9Hz,3H),HRESIMS m/z＝771.2446[M+H]+(calcd for C37H42N2O14S,770.2357).
EXAMPLE 10 preparation of (R) -14-O- {5- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -5-oxovaleryl } -triptolide (LA-59)
According to the preparation method of example 2, it is prepared from triptolide and intermediate 2j with a yield of 68%. H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.64-7.60(t,2H),5.06(s,1H),4.66(s,3H),4.46-4.43(m,2H),3.81-3.80(d,J＝3.0Hz,1H),3.51-3.50(d,J＝2.3Hz,1H),3.45-3.44(d,J＝5.6Hz,1H),1.31-1.30(d,J＝6.4Hz,3H),0.93-0.92(d,J＝6.9Hz,3H),0.87-0.85(d,J＝6.9Hz,1H),0.82-0.80(d,J＝6.9Hz,3H),HRESIMS m/z＝771.2446[M+H]+(calcd for C37H42N2O14S,770.2357).
EXAMPLE 11 preparation of (S) -14-O- {5- [ (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -3-butoxy ] -5-oxovaleryl } -triptolide (LA-60)
According to the preparation method of example 2, the product is prepared from triptolide and intermediate 2k with a yield of 65%. H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.64-7.60(t,2H),5.06(s,1H),4.66(s,3H),4.46-4.43(m,2H),3.81-3.80(d,J＝3.0Hz,1H),3.51-3.50(d,J＝2.3Hz,1H),3.45-3.44(d,J＝5.6Hz,1H),1.31-1.30(d,J＝6.4Hz,3H),0.93-0.92(d,J＝6.9Hz,3H),0.87-0.85(d,J＝6.9Hz,1H),0.82-0.80(d,J＝6.9Hz,3H),HRESIMS m/z＝771.2446[M+H]+(calcd for C37H42N2O14S,770.2357).
EXAMPLE 12 preparation of 14-O- {5- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -butoxy ] -5-oxovaleryl } -triptolide (LA-61)
According to the preparation method of example 2, it was prepared from triptolide and intermediate 2l in a yield of 68%. H1NMR(400MHz,CDCl3)δ8.07-8.05(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.64-7.60(t,2H),5.09(s,1H),4.66(m,2H),4.47-4.45(t,2H),4.18-4.16(t,3H),4.09-4.08(d,J＝6.8Hz,1H),3.82(d,J＝3.0Hz,1H),3.53-3.52(d,J＝2.3Hz,1H),3.47-3.46(d,J＝5.6Hz,1H),1.04(s,3H),0.99-0.98(d,J＝6.9Hz,3H),0.96(J＝6.9Hz,3H),0.85-0.84(d,6.9Hz,3H).HRESIMS m/z＝771.2446[M+H]+(calcd for C37H42N2O14S,770.2357).
EXAMPLE 13 preparation 114-O- {4- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -2-yne-butoxy ] -5-oxovaleryl } -triptolide (LA-62)
According to the preparation method of example 2, the preparation was prepared from triptolide and intermediate 2m with a yield of 64%. H1NMR(400MHz,CDCl3)δ8.09-8.07(d,J＝7.8Hz,2H),7.76-7.73(t,1H),7.63-7.60(t,2H),5.15(s,2H),4.76(s,2H),4.66(s,2H),3.84(d,J＝3.0Hz,1H),3.56(d,J＝2.3Hz,1H),3.50-3.49(d,J＝5.6Hz,1H),1.58-1.54(m,1H),1.24(s,2H),1.04(s,3H),0.95-0.93(d,J＝6.9Hz,3H),0.84-0.82(d,J＝6.9Hz,3H).HRESIMS m/z＝767.2112[M+H]+(calcd for C37H38N2O14S,766.2043).
EXAMPLE 14 preparation of 14-O- {4- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) ] -butyryl } -triptolide (LA-63)
According to the preparation method of example 2, it is prepared from triptolide and intermediate 4a with a yield of 77%.
H1NMR(400MHz,CDCl3)δ8.05-8.04(d,J＝7.8Hz,2H),7.77-7.74(t,1H),7.65-7.62(t,2H),5.08(s,1H),4.70-4.62(m,2H),4.53-4.51(t,2H),3.82(d,J＝3.0Hz,1H),3.53(d,J＝2.3Hz,1H),3.46-3.45(d,J＝5.6Hz,1H),1.23(s,3H),0.99(s,3H),0.95-0.94(d,J＝6.9Hz,3H),0.83-0.82(d,6.9Hz,3H).HRESIMS m/z＝671.1928[M+H]+(calcd for C32H34N2O12S,670.1905).
EXAMPLE 15 preparation of 14-O- {4- [4- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) ] -butoxyacyl } -triptolide (LA-64)
According to the preparation method of example 2, the compound is prepared from triptolide and intermediate 4b, and the yield is 72%.
H1NMR(400MHz,CDCl3)δ8.09-8.08(d,J＝7.8Hz,2H),7.76-7.73(t,1H),7.63-7.60(t,2H),5.15(s,1H),4.39-4.36(d,J＝16.7Hz,1H)4.29-4.26(d,J＝16.7Hz,1H),3.84(d,J＝3.0Hz,1H),3.56(d,J＝2.3Hz,1H),3.50-3.49(d,J＝5.6Hz,1H),1.23(s,3H),0.99(s,3H),0.95-0.94(d,J＝6.9Hz,3H),0.83-0.82(d,6.9Hz,3H).HRESIMS m/z＝687.188[M+H]+(calcd for C32H34N2O13S,686.1854).
EXAMPLE 16 preparation of 14-O- {4- [5- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) ] -pentanoyl } -triptolide (LA-65)
According to the preparation method of example 2, it is prepared from triptolide and intermediate 4c in 81% yield.
H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.76-7.73(t,1H),7.63-7.60(t,2H),5.09(s,1H),4.70-4.62(m,2H),4.47-4.44(t,2H),3.83-3.82(d,J＝3.0Hz,1H),3.47-3.46(d,J＝5.6Hz,1H),1.23(s,3H),1.03(s,3H),0.96-0.95(d,J＝6.9Hz,3H),0.84-0.83(d,6.9Hz,3H).HRESIMS m/z＝685.2081[M+H]+(calcd for C33H36N2O12S,684.2062).
EXAMPLE 17 preparation of 14-O- {4- [5- (2-oxo-3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) ] -hexanoyl } -triptolide (LA-66)
According to the preparation method of example 2, the triptolide is prepared from triptolide and the intermediate 4d, and the yield is 75%.
H1NMR(400MHz,CDCl3)δ8.06-8.04(d,J＝7.8Hz,2H),7.78-7.74(t,1H),7.65-7.61(t,2H),5.09(s,1H),4.70-4.62(m,2H),4.52(s,1H),4.44-4.41(t,2H),3.83(d,J＝3.0Hz,1H),3.54(d,J＝2.5Hz,1H),3.46-3.45(d,J＝5.6Hz,1H),1.04(s,3H),0.97-0.95(s,3H),0.85-0.83(s,3H).HRESIMS m/z＝699.2219[M+H]+(calcd for C34H38N2O12S,699.2218).
EXAMPLE 18 preparation of 14-O- (3-benzenesulfonyl-1, 2, 5-oxadiazol-4-oxo) -triptolide (LA-67)
500mg of triptolide was completely dissolved in 10ml of tetrahydrofuran. The ice-water bath was cooled to 0 ℃ and 111mg of sodium hydride was slowly added thereto in portions and stirred for 10 minutes. Then, a tetrahydrofuran solution (1.02g in 10ml of tetrahydrofuran) of benzenesulfonyl furazan nitroxide was dropped into the reaction flask, and stirring was continued for 2 hours. Stopping reaction, adding dichloromethane for dilution, washing with a small amount of water and saturated sodium chloride solution once respectively, drying with anhydrous sodium sulfate, filtering, and spin-drying the filtrate to obtain a crude product. Purification was performed by silica gel column chromatography (chloroform: methanol 20:1) to obtain a white solid, and recrystallization was performed with methanol to obtain 511mg of a pure product, with a yield of 63%. H1NMR(400MHz,CDCl3)δ8.23-8.21(d,J＝7.8Hz,2H),7.75-7.72(t,1H),7.62-7.59(t,2H),5.05(s,1H),4.72-4.63(q,2H),3.95-3.94(d,J＝2.2Hz,1H),3.69(s,1H),3.48-3.47(d,J＝5.4Hz,1H),2.75-2.72(m,1H),2.39-2.35(m,1H),2.22-2.17(m,2H),2.10-2.05(m,1H),1.91-1.85(t,1H),1.68-1.65(m,1H),1.31-1.25(m,1H),1.09(s,3H),1.04-1.02(d,J＝6.80Hz,3H),0.94-0.93(d,J＝6.80Hz,3H)。HRESIMS m/z＝585.1521[M+H]+(calcd for C28H28O10N2S,584.1465)。
Pharmacological experiments
Experimental example 1 cytotoxic Activity test
1. Experimental materials: tetramethyl azoazolium salt (MTT, produced by Serva, prepared temporarily with fresh 1640 culture solution), BGC-823 (human gastric cancer cells), HCT-116 (human colon cancer cells), NCI-H1650 (human lung cancer cells), HepG2 (human liver cancer cells), BIORAD 550 enzyme-linked immunosorbent assay.
2. Experimental methods
(1) And (3) cell culture: cells were cultured in RRMI1640 medium containing 10% fetal bovine serum, 100U/mL penicillin and 100mg/L at 37 ℃ in 5% CO2Subculturing in a saturated humidity incubator, and selecting logarithmic growth cycle cells for experiments.
(2) MTT method: taking cells in logarithmic growth phase, digesting, fully blowing and beating into single cell suspension, counting, diluting into 1 × 104cells/mL, seeded in 96-well plates at 100. mu.L/well. 4-5 concentration levels were set for each sample, and then 100. mu.L of medium from samples of different concentration levels, each concentration level being 3 wells in parallel, was added to the experimental wells. The control group was added with an equal volume of solvent. Placing 96-well culture plate at 37 deg.C and 5% CO2After culturing for 96 hours in a saturated humidity incubator, the culture medium was discarded, a freshly prepared serum-free medium containing 0.20mg/mL MTT was added to each well, and after further culturing for 4 hours at 37 ℃, the supernatant was removed by centrifugation. Add 150. mu.L DMSO per well to dissolve formazan precipitate, and place for 5 minutes with micro-shaking to dissolve it sufficiently. The optical density at 570nm was determined on a BIORAD 550 model enzyme calibrator. Calculating the tumor cell growth inhibition rate according to the following formula, plotting the drug concentration against the tumor cell growth inhibition rate to obtain a measurement curve, and reading the half Inhibition Concentration (IC) of the drug from the curve50) The value is obtained.
Tumor cell growth inhibition (%) was (1-assay/control wells) × 100%
3. The experimental results are as follows: as shown in table 1.
TABLE 1 tumor cytotoxic Activity
Figure BDA0001661798100000121
Experimental example 2 in vivo antitumor Effect of LA-67 against mouse melanoma B16F10
1. Experimental materials: a test compound LA-67; cyclophosphamide (CTX, positive drug); c57 mice, 18-20g in weight, SPF grade, female, were provided by the institute of medical laboratory animals, national academy of medical sciences.
2. The experimental method comprises the following steps:
(1) and the step: grinding subcultured B16F10 tumor with homogenizer, washing with sterile normal saline twice, counting, adjusting cell concentration to 25 × 10 with normal saline6Perml, 0.2ml of the cell suspension was adjusted to inoculate the right underarm of the mice. Animals were randomly grouped the next day after inoculation, 7 animals per group, dosed after weighing, the compound to be tested was dosed 1 time per day, dosed 20 times in succession, weighed, animals sacrificed, tumor tissue stripped and weighed. And finally, calculating the tumor inhibition rate, and evaluating the anti-tumor effect intensity by using the tumor inhibition rate.
(2) Grouping: control group (Control), positive drug Control group (CTX)60mg/kg (abdominal cavity); LA-670.3 mg/kg (gavage), LA-671 mg/kg (gavage).
(3) And the calculation method comprises the following steps:
relative tumor proliferation rate T/C (%): T/C% ═ T/C × 100%. (T: tumor weight in treatment group; C: tumor weight in negative control group).
Tumor proliferation inhibition ratio TGI (%): TGI ═ 1-T/C) × 100. (T: tumor weight in treatment group; C: tumor weight in negative control group).
3. The experimental results are as follows:
treatment animals were weighed after 20 days of administration and the effect of the compound on mouse B16F10 is shown in figure 1 and table 2.
TABLE 2 in vivo antitumor Effect of Compounds on mouse B16F10
Figure BDA0001661798100000131
1.TTEST,*P<0.05,**P<0.01
NA not applicable.
Experimental example 3 preliminary oral acute toxicity test of LA-67
1. Experimental materials: a test compound LA-67; a mouse weightometer; gavage needle (No. 12); 50 ICR mice, each half of male and female, weighing 18-20g, were purchased from Beijing Huafukang Biotech GmbH.
2. The experimental method comprises the following steps: 50 ICR mice were randomly divided into five groups, 100mg/kg LA-67, 125mg/kg LA-67, 150mg/kg LA-67, 175mg/kg LA-67, 200mg/kg LA-67. Test compounds were administered by gavage 1 time and survival was recorded after 14 days.
3. The experimental results are as follows: preliminary oral acute toxicity experiments show LD of LA-6750160.85mg/kg, the toxicity is much less than triptolide. Table 3 shows the survival statistics for each group of animals, and Table 4 shows the LD calculated by the DASver 1.0 software according to the Bliss method50。
TABLE 3 survival rates of the groups of animals
Figure BDA0001661798100000141
TABLE 4LD50 calculation results
Figure BDA0001661798100000142

Claims (7)

1. A compound having the structure of formula I or a pharmaceutically acceptable salt thereof,
Figure FDA0003358506930000011
R1is selected from (CH)2)n、C6H4CH or CH2C(CH3)2CH2；
R2Is selected from (CH)2)n、CH(CH3)CH2CH2、H2CC≡CCH2Or (CH)2)2O(CH2)2；
Wherein n is 1,2, 3, 4, 5, 6, 7 or 8;
ph represents a phenyl group.
2. A compound having the structure of formula II or a pharmaceutically acceptable salt thereof,
Figure FDA0003358506930000012
R3is selected from (CH)2)n、CH2O(CH2)2、C≡CCH2；
Wherein n is 1,2, 3, 4, 5, 6, 7 or 8;
ph represents a phenyl group.
3. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:
Figure FDA0003358506930000013
LA-50 R1＝CH2CH2 R2＝CH(CH3)CH2CH2 LA-51 R1＝CH2CH2 R2＝(R)-CH(CH3)CH2CH2
LA-52 R1＝CH2CH2 R2＝(S)-CH(CH3)CH2CH2 LA-53 R1＝CH2CH2 R2＝CH2CH2CH2CH2
LA-54 R1＝CH2CH2 R2＝H2CC≡CCH2 LA-55 R1＝CH2CH2 R2＝(CH2)2O(CH2)2
LA-56 R1＝CH2CH2 R2＝CH2CH2CH2 LA-57 R1＝CH2CH2 R2＝(CH2)5
LA-58 R1＝(CH2)3 R2＝CH(CH3)CH2CH2 LA-59 R1＝(CH2)3 R2＝(R)-CH(CH3)CH2CH2
LA-60 R1＝(CH2)3 R2＝(S)-CH(CH3)CH2CH2 LA-61 R1＝(CH2)3 R2＝CH2CH2CH2CH2
LA-62 R1＝(CH2)3 R2＝H2CC≡CCH2。
4. a compound according to claim 2, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:
Figure FDA0003358506930000021
LA-63 R3＝CH2CH2CH2 LA-64 R3＝CH2O(CH2)2
LA-65 R3＝(CH2)4 LA-66 R3＝(CH2)5
wherein Ph represents a phenyl group.
5. A compound represented by the formula or a pharmaceutically acceptable salt thereof, said compound selected from the group consisting of:
Figure FDA0003358506930000022
wherein Ph represents a phenyl group.
6. A pharmaceutical composition comprising the compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier or excipient.
7. Use of a compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a neoplastic disease.
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CN102153610A (en) * 2011-02-28 2011-08-17 中国药科大学 Nitric oxide donor type bile acid derivatives as well as preparation methods and medicinal application thereof
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