CN102101864B - Trigonolactone A-D and medicinal composition, preparation method and application thereof - Google Patents
Trigonolactone A-D and medicinal composition, preparation method and application thereof Download PDFInfo
- Publication number
- CN102101864B CN102101864B CN200910218391A CN200910218391A CN102101864B CN 102101864 B CN102101864 B CN 102101864B CN 200910218391 A CN200910218391 A CN 200910218391A CN 200910218391 A CN200910218391 A CN 200910218391A CN 102101864 B CN102101864 B CN 102101864B
- Authority
- CN
- China
- Prior art keywords
- trigonostemon
- compound
- trigonostemon chinensis
- formula
- preparation
- 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.)
- Active
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides Trigonolactone A-D shown as a structural formula (I). The Trigonolactone A-D is macrocyclic diterpenoids with a novel chemical structure has remarkable HIV-1 (Human Immunodeficiency Virus 1) induction C8166 cell formation synplasm suppression activity, has the therapeutic index (TI) of between 1848 and 13484, and can be taken as an anti-human immunodeficiency virus (HIV) drug. In Trigonostelliimone A(1), R1 is Cl; and R2 is H; and in HTrigonostelliimone B(2), R1 is H and R2 is H.
Description
Technical field:
The invention belongs to technical field of pharmaceuticals, particularly, relate to Trigonostemon chinensis lactone A-D, it is the pharmaceutical composition of active constituents of medicine, its preparation method, its application in the preparation anti-AIDS drug.
Background technology: Trigonostemon (Trigonostemon) China has 6 kinds, mainly is distributed in China south China and southwest.Up to now, do not see in the prior art that Trigonostemon chinensis lactone A-D compound and active report thereof are arranged.
Summary of the invention:
The object of the present invention is to provide the Trigonostemon chinensis lactone A-D shown in the structural formula (I); Anti-AIDS pharmaceutical composition and medicine that it is formed for active constituents of medicine; Its preparation method, and this compounds and the application of pharmaceutical composition in preparation treatment AIDS-treating medicine thereof.
Above-mentioned purpose of the present invention is to be achieved through following technical scheme:
The preparation method of formula (I) compound gets the Meng Lun Trigonostemon chinensis, extracts three times with 95% alcohol heating reflux; Each 4 hours, united extraction liquid, decompression recycling ethanol; Get medicinal extract, its water is pinched molten, filtration, use sherwood oil respectively; Ethyl acetate extraction three times; Get ethyl acetate extraction part, get Fr1, Fr2, Fr3, Fr4 four parts, wherein sherwood oil with sherwood oil-acetone gradient elution: 7: 3 wash-out parts of acetone Fr3 must contain the component of Trigonostemon chinensis lactone A-D with Sephadex LH-20 methanol-eluted fractions; This component recycle silicon plastic column chromatography chloroform: 8: 1 wash-outs of methyl alcohol, use the HPLC acetonitrile then: 65: 35 purifying of water get compound Trigonostemon chinensis lactone A, B, C and D.
It is Trigonolactone A-D (1-4) that the present invention provides the Trigonostemon chinensis lactone A-D compound shown in the structural formula (I),
Trigonosteliimone?A(1)R
1＝Cl R
2＝H
Trigonosteliimone?B(2)R
1＝H R
2＝H
Trigonosteliimone?C(3)R
1＝Cl
Trigonosteliimone?D(4)R
1＝H
The inverase compsn contains compound and the pharmaceutically acceptable carrier shown in claim 1 formula (I) of treating significant quantity.
The application of formula (I) compound in the preparation anti-AIDS drug.
Contain the application of pharmaceutical composition in the preparation anti-AIDS drug that formula (I) compound of treating significant quantity and pharmaceutically acceptable carrier are formed.
When The compounds of this invention is used as medicine, can directly use, perhaps use with the form of pharmaceutical composition.This pharmaceutical composition contains 0.1-99%, is preferably the The compounds of this invention of 0.5-90%, and all the other are acceptable on the pharmacology, nontoxic and inert pharmaceutically acceptable carrier and/or vehicle to humans and animals.
Described pharmaceutical carrier or vehicle are one or more solids, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.The method that the compsn of orcinol-1-oxygen-β-D-glucose pyrans glycosides, orcinol and verivate thereof adopts pharmacy and field of food to generally acknowledge is prepared into various formulations, like liquid preparation (injection, suspensoid, emulsion, solution, syrup etc.), solid preparation (tablet, capsule, granule, electuary etc.), spray, aerosol etc.Medicine of the present invention can carry out the treatment of anti-AIDS through route of administration such as injection (intravenous injection, intravenous drip, intramuscular injection, abdominal injection, subcutaneous injection) and oral, sublingual administration, mucous membrane dialysis.
Embodiment:
Further specify essentiality content of the present invention with the embodiment of the invention below, but do not limit the present invention with this.
Embodiment 1:
Meng Lun Trigonostemon chinensis (Trigonostemon lii Y.T.Chang) complete stool 20kg, with 95% alcohol heating reflux extract three times (3 * 4h), united extraction liquid, decompression recycling ethanol, medicinal extract 2kg.Its water is pinched molten, filtration, use sherwood oil respectively, ethyl acetate extraction three times obtains ethyl acetate extraction part 200g; Get Fr1, Fr2, Fr3, Fr4 four parts, wherein sherwood oil with sherwood oil-acetone gradient elution: 7: 3 wash-out parts of acetone Fr3 must contain the component of Trigonostemon chinensis lactone A-D with the SephadexLH-20 methanol-eluted fractions; This component recycle silicon plastic column chromatography chloroform: 8: 1 wash-outs of methyl alcohol, use the HPLC acetonitrile then: 65: 35 purifying of water get compound Trigonostemon chinensis lactone A (Trigonosteliimone A) (1) (50.4mg), B (2) (12.0mg), C (3) (35.5mg) and D (4) (45.8mg).
4 compound Trigonosteliimone A (1) that separation obtains), the structural analysis data of B (2), C (3) and D (4) are following:
Trigonostemon chinensis lactone A (Trigonosteliimone A) (1): white amorphous powder; C
38H
49O
11Cl;
(c 0.20 CHCl
3); UV (CHCl
3) λ
Max(257.6nm ε 17474); IR (KBr) v
Max3455,2931,1713 and 1618cm
-1ESIMSm/z 739 [M+Na]
+HRESIMS m/z739.2866 [M+Na]
+(calcd forC
38H
49O
11NaCl, 739.2861).
1H NMR (CDCl
3, 400 MHz) and δ: 6.48 (1H, s, H-1), 5.38 (1H, s, H-3), 3.93 (1H, s, H-5), 3.41 (1H, s, H-7); 4.74 (1H, s, H-8), 2.74 (1H, q, J=8.8Hz, H-11), 4.29 (1H, s, H-12), 4.39 (1H, s, H-14); (1.47 1H, m, H-16 α), 1.73 (1H, m, H-16 β), 1.39 (3H, s, H-17), 1.23 (3H, d, J=8.5Hz, H-18); 6.07 (1H, s, H-19), 3.83 (2H, br s, H-20), 2.41 (1H, m, H-2 '), 1.70 (1H, m, H-3 α '), 1.36 (1H; M, H-3 β '), 1.35 (1H, m, H-4 α '), 1.09 (1H, m, H-4 β '), 1.31 (2H, m, H-5 '), 1.41 (2H, m; H-6 '), 1.48 (1H, m, H-7 α '), 1.18 (1H, m, H-7 β '), 1.23 (1H, m, H-8 α '), 1.40 (1H, m, H-8 β '); (1.59 1H, m, H-9 '), 1.04 (3H, d, J=8.0Hz, H-10 '), 1.20 (3H, d, J=9.0Hz, H-11 '), 7.76 (1H, m; H-3 "), 7.43 (1H, m, H-4 "), 7.41 (1H, m, H-5 "), 7.43 (1H, m, H-6 "), 7.76 (1H, m, H-7 ").
13C NMR (CDCl
3, 100.6MHz) δ: 125.7 (C-1, d), 139.7 (C-2, s), 78.6 (C-3, d), 83.4 (C-4, s), 7 1.6 (C-5, d); 60.5 (C-6, s), 62.3 (C-7, d), 34.6 (C-8, d), 74.6 (C-9, s), 149.2 (C-10, s), 34.7 (C-11; D), 79.6 (C-12, d), 72.1 (C-13, s), 79.2 (C-14, d), 75.5 (C-15, s), 38.3 (C-16, t); 23.7 (C-17, q), 13.9 (C-18, q), 114.5 (C-19, d), 63.6 (C-20, t), 178.7 (C-1 ', s), 42.3 (C-2 '; D), 34.9 (C-3 ', t), 31.4 (C-4 ', t), 27.0 (C-5 ', t), 28.2 (C-6 ', t), 27.8 (C-7 ', t); 37.8 (C-8 ', t), 25.1 (C-9 ', d), 24.9 (C-10 ', q), 18.6 (C-11 ', q), 108.4 (C-1 ", s), 138.2 (C-2 "; S), 125.2 (C-3 ", d), 128.2 (C-4 ", d), 129.5 (C-5 ", d), 128.2 (C-6 ", d), 125.2 (C-7 ", d).
Trigonostemon chinensis lactone B (Trigonosteliimone B) (2): white amorphous powder; C
38H
50O
11 
(c 0.14 CHCl
3); UV (CHCl
3) λ
Max(246.6nm ε 15144); IR (KBr) v
Max3442,2924,1710 and 1639cm
-1ESIMS m/z683 [M+H]
+HRESIMS m/z683.3441 [M+H]
+(calcd for C
38H
51O
11, 683.3431).
1H NMR (CDCl
3, 400 MHz) and δ: 6.30 (1H, s, H-1), 5.38 (1H, s, H-3), 3.87 (1H, s, H-5), 3.49 (1H, s, H-7), 4.79 (1H; S, H-8), 2.77 (1H, q, J=9.0Hz, H-11), 4.29 (1H, s, H-12), 4.39 (1H, s, H-14), 1.43 (1H, m; H-16 α), 1.71 (1H, m, H-16 β), 1.37 (3H, s, H-17), 1.23 (3H, d, J=8.5Hz, H-18), 5.17 (1H, s, H-19a); 5.07 (1H, s, H-19b), 3.75 (1H, d, J=15.0Hz, H-20 α), 3.91 (1H, d, J=15.0Hz, H-20 β), 2.35 (1H, m, H-2 '); (1.69 1H, m, H-3 α '), 1.30 (1H, m, H-3 β '), 1.30 (1H, m, H-4 α '), 1.06 (1H, m, H-4 β '), 1.30 (2H, m; H-5 '), 1.43 (2H, m, H-6 '), 1.48 (1H, m, H-7 α '), 1.15 (1H, m, H-7 β '), 1.21 (1H, m, H-8 α '), 1.36 (1H; M, H-8 β '), 1.55 (1H, m, H-9 '), 1.04 (3H, d, J=8.0Hz, H-10 '), 1.19 (3H, d, J=9.5 Hz, H-11 '), 7.76 (1H; M, H-3 "), 7.41 (1H, m, H-4 "), 7.41 (1H, m, H-5 "), 7.41 (1H, m, H-6 "), 7.76 (1H, m, H-7 ").
13C NMR (CDCl
3, 100.6 MHz) and δ: 130.2 (C-1, d), 145.6 (C-2, s), 80.0 (C-3, d), 83.5 (C-4, s), 71.8 (C-5, d); 60.0 (C-6, s), 62.0 (C-7, d), 34.5 (C-8, d), 74.5 (C-9, s), 147.5 (C-10, s), 34.6 (C-11; D), 79.7 (C-12, d), 72.1 (C-13, s), 79.4 (C-14, d), 75.3 (C-15, s), 37.9 (C-16, t); 23.8 (C-17, q), 13.9 (C-18, q), 111.2 (C-19, t), 62.4 (C-20, t), 179.7 (C-1 ', s), 42.8 (C-2 '; D), 35.1 (C-3 ', t), 31.6 (C-4 ', t), 27.8 (C-5 ', t), 28.3 (C-6 ', t), 27.2 (C-7 ', t); 37.5 (C-8 ', t), 24.9 (C-9 ', d), 25.0 (C-10 ', q), 18.8 (C-11 ', q), 108.4 (C-1 ", s), 138.5 (C-2 "; S), 125.2 (C-3 ", d), 128.2 (C-4 ", d), 129.5 (C-5 ", d), 128.2 (C-6 ", d), 125.2 (C-7 ", d).
Trigonostemon chinensis lactone C (Trigonosteliimone C) (3): white amorphous powder; C
45H
53O
14Cl;
(c 0.67 CHCl
3); UV (CHCl
3) λ
Max(248.4nm ε 20467); IR (KBr) v
Max3444,2925,1710 and 1677cm
-1ESIMSm/z 853 [M+H]
+HRESIMS m/z853.3245 [M+H]
+(calcd forC
45H
54O
14Cl, 853.3202).
1H NMR (CDCl
3, 400MHz) δ: 6.42 (1H, s, H-1), 5.32 (1H, s, H-3), 3.78 (1H, s, H-5), 2.99 (1H, s, H-7), 4.45 (1H; S, H-8), 2.71 (1H, q, J=8.5Hz, H-11), 4.31 (1H, s, H-12), 4.48 (1H, s, H-14), 1.84 (1H, d; J=18.0Hz, H-16 α), 1.56 (1H, dd, J=7.5,18.0Hz, H-16 β), 4.80 (1H, d, J=15.0Hz, H-17 α), 4.57 (1H, d, J=15.0Hz; H-17 β), 1.24 (3H, d, J=9.0Hz, H-18), 6.00 (1H, s, H-19), 3.59 (1H, d, J=15.5Hz, H-20 α), 3.48 (1H, d; J=15.5Hz, H-20 β), 2.31 (1H, m, H-2 '), 1.67 (1H, m, H-3 α '), 1.30 (1H, m, H-3 β '), 1.31 (1H, m; H-4 α '), 1.12 (1H, m, H-4 β '), 1.55 (2H, m, H-5 '), 1.13 (2H, m, H-6 '), 1.54 (1H, m, H-7 α '); (1.16 1H, m, H-7 β '), 1.40 (1H, m, H-8 α '), 1.30 (1H, m, H-8 β '), 1.76 (1H, m, H-9 '), 1.09 (3H; D, J=8.0Hz, H-10 '), 1.17 (3H, d, J=9.0Hz, H-11 '), 7.76 (1H, m, H-3 "), 7.43 (1H, m, H-4 "); 7.41 (1H, m, H-5 "), 7.43 (1H, m, H-6 "), 7.76 (1H, m, H-7 "), 7.18 (1H, d, J=10.0Hz, H-4 " '); 7.49 (1H, t, J=10.0Hz, H-5 " '), 6.92 (1H, t, J=10.0Hz, H-6 " '), 7.70 (1H, dd, J=1.5,10.0Hz, H-7 " ').
13C NMR (CDCl
3, 100.6MHz) δ: 125.7 (C-1, d), 139.6 (C-2, s), 78.6 (C-3, d), 83.8 (C-4, s), 71.0 (C-5, d), 60.3 (C-6; S), 61.8 (C-7, d), 34.6 (C-8, d), 74.6 (C-9, s), 149.0 (C-10, s), 34.7 (C-11, d), 79.2 (C-12; D), 72.3 (C-13, s), 79.2 (C-14, d), 76.0 (C-15, s), 37.1 (C-16, t), 67.8 (C-17, t), 13.9 (C-18; Q), 114.5 (C-19, d), 63.4 (C-20, t), 178.8 (C-1 ', s), 42.5 (C-2 ', d), 34.7 (C-3 ', t); 31.0 (C-4 ', t), 28.0 (C-5 ', t), 28.4 (C-6 ', t), 27.1 (C-7 ', t), 37.9 (C-8 ', t), 25.6 (C-9 '; D), 24.2 (C-10 ', q), 18.8 (C-11 ', q), 108.4 (C-1 ", s), 138.4 (C-2 ", s), 125.2 (C-3 ", d); 128.2 (C-4 ", d), 129.5 (C-5 ", d), 128.2 (C-6 ", d), 125.2 (C-7 ", d), 170.2 (C-1 " ', s), 112.2 (C-2 " '; s), 161.7 (C-3 " ', s), 118.2 (C-4 " ', d), 136.1 (C-5 " ', d), 119.0 (C-6 " ', d), 129.5 (C-7 " ', d).
Trigonostemon chinensis lactone D (Trigonosteliimone D) (4): white amorphous powder; C
45H
54O
14 
(c 0.63 CHCl
3); UV (CHCl
3) λ
Max(244.2nm ε 28282); IR (KBr) v
Max3443,2925,1704 and 1678cm
-1ESIMS m/z841 [M+Na]
+HRESIMS m/z 841.3407 [M+Na]
+(calcd forC
45H
54O
14Na, 841.3411).
1H NMR (CDCl
3, 400 MHz) and δ: 6.12 (1H, s, H-1), 5.35 (1H, s, H-3), 3.78 (1H, s, H-5), 3.01 (1H, s, H-7), 4.46 (1H; S, H-8), 2.68 (1H, q, J=7.2 Hz, H-11), 4.29 (1H, s, H-12), 4.43 (1H, s, H-14), 1.87 (1H, d; J=18.0 Hz, H-16 α), 1.56 (1H, dd, J=7.5,18.0Hz, H-16 β), 4.78 (1H, d, J=11.6Hz, H-17 α), 4.54 (1H, d, J=11.6Hz; H-17 β), 1.24 (3H, d, J=9.0Hz, H-18), 5.11 (1H, s, H-19a), 6.01 (1H, s, H-19b), 3.56 (1H, br s, H-20 α); (3.42 1H, br s, H-20 β), 2.31 (1H, m, H-2 '), 1.67 (1H, m, H-3 α '), 1.28 (1H, m, H-3 β '), 1.31 (1H, m; H-4 α '), 1.11 (1H, m, H-4 β '), 1.52 (2H, m, H-5 '), 1.15 (2H, m, H-6 '), 1.52 (1H, m, H-7 α '); (1.16 1H, m, H-7 β '), 1.40 (1H, m, H-8 α '), 1.29 (1H, m, H-8 β '), 1.73 (1H, m, H-9 '), 1.07 (3H; D, J=7.5Hz, H-10 '), 1.18 (3H, d, J=9.0Hz, H-11 '), 7.76 (1H, m, H-3 "), 7.43 (1H, m, H-4 "); 7.41 (1H, m, H-5 "), 7.43 (1H, m, H-6 "), 7.76 (1H, m, H-7 "), 7.18 (1H, d, J=10.0 Hz, H-4 " '); 7.49 (1H, t, J=10.0Hz, H-5 " '), 6.92 (1H, t, J=10.0Hz, H-6 " '), 7.70 (1H, dd, J=1.5,10.0Hz, H-7 " ').
13C NMR (CDCl
3, 100.6MHz) δ: 130.2 (C-1, d), 145.6 (C-2, s), 79.2 (C-3, d), 83.2 (C-4, s), 71.2 (C-5, d), 60.1 (C-6; S), 61.8 (C-7, d), 34.2 (C-8, d), 74.5 (C-9, s), 147.2 (C-10, s), 34.6 (C-11, d), 79.9 (C-12; D), 72.4 (C-13, s), 79.3 (C-14, d), 75.8 (C-15, s), 37.2 (C-16, t), 67.8 (C-17, t), 13.9 (C-18; Q), 111.3 (C-19, t), 62.8 (C-20, t), 179.2 (C-1 ', s), 42.5 (C-2 ', d), 34.9 (C-3 ', t); 31.3 (C-4 ', t), 28.2 (C-5 ', t), 28.3 (C-6 ', t), 27.2 (C-7 ', t), 37.9 (C-8 ', t), 25.4 (C-9 '; D), 24.1 (C-10 ', q), 18.8 (C-11 ', q), 108.4 (C-1 ", s), 138.4 (C-2 ", s), 125.2 (C-3 ", d); 128.2 (C-4 ", d), 129.5 (C-5 ", d), 128.2 (C-6 ", d), 125.2 (C-7 ", d), 170.2 (C-1 " ', s), 112.2 (C-2 " '; s), 161.8 (C-3 " ', s), 118.2 (C-4 " ', d), 135.9 (C-5 " ', d), 118.9 (C-6 " ', d), 129.4 (C-7 " ', d).
Embodiment 2:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, by the used assistant agent of conventional injection liquid, adds the injection water respectively, smart filter, and injection liquid is processed in the embedding sterilization.
Embodiment 3:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, and it is dissolved in respectively in the sterile water for injection, and stirring makes molten, filters with aseptic suction funnel, and aseptic more smart filter is sub-packed in 2 ampoules, and aseptic sealing by fusing gets powder injection behind the frozen drying.
Embodiment 4:
The Trigonostemon chinensis lactone A-D (1-4) that obtains is that 9: 1 ratio adds vehicle with the vehicle weight ratio respectively with separate, and processes pulvis.
Embodiment 5:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, is 1 in itself and vehicle weight ratio respectively: 5-1: 10 ratio adds vehicle, pelletizing press sheet.
Embodiment 6:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, processes oral liquid by conventional oral liquid method for making respectively.
Embodiment 7:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, is that 5: 1 ratio adds vehicle in itself and vehicle weight ratio respectively, processes capsule or granule or electuary.
Embodiment 8:
Method by embodiment 1 makes Trigonostemon chinensis lactone A-D (1-4) earlier, is that 3: 1 ratio adds vehicle in itself and vehicle weight ratio respectively, processes capsule or granule or electuary.
The novel macrocyclic diterpene lactone of above-mentioned 4 chemical structures of the present invention's isolation identification from the Meng Lun Trigonostemon chinensis (Trigonostemon.liiY.T.Chang) that originates from Xishuangbanna, Yunnan
(1-4); Through the HIV-resistant activity screening, find that this compounds has significant HIV-1 and induces the C8166 cell to form synplasm inhibition activity, with reference to its cytotoxicity to C8166; Therapeutic index (TI) is a kind of strong HIV-resistant activity medicine that has in the 1848-13484 scope.
The pharmacological tests of The compounds of this invention is described with Test Example of the present invention below, and it is strong active to be used to prove that The compounds of this invention has AIDS.
Test Example 1:
The anti-HIV pharmacological action of Trigonostemon chinensis lactone A-D (1-4) experimental result:
I material and method
One, measures medicine and compound
Testing sample is by 4 compd A-D of formula provided by the invention (I) (1-4).Testing sample is dissolved among the DMSO, 4 ℃ of preservations, and storage concentration is 25.0mg/ml;
Two, reagent and solution
(1) reagent
HEPES (N-2 (2-Hydroxyothyl) piperazine-N '-(2-ethanesufonicacid), MTT (3; (4; 5-dimethylthiazol-2-yl)-2; 5-diphenyl tetrazoliumbromide), DMF (N, N '-Dimethyl formamine), penicillium mould (Penicillin), Vetstrep (Streptomycin sulfate), Stimulina (Glutamine) are all available from Sigma company; (2-ME 2-Mercaptoethanol) is the Bio-Rad Company products to 2 mercapto ethanol.RPMI-1640 and newborn calf serum are the Gibco Company products.
(2) substratum
The RPMI-1640 perfect medium contains 10% newborn calf serum, 2mM L-glutaminate, 10mM HEPES, 50 μ M 2 mercapto ethanols, 100,000IU penicillium mould, 100 μ g/ml Streptomycin sulphates.
Three, cell and virus
The human T lymphocyte is that C8166, MT-4 and HIV-1 test strain HIV-1IIIB by Britain Medical Research Council, and AIDS Reagent Project is so kind as to give.All cells is all cultivated with the RPMI-1640 perfect medium that contains 10% calf serum with virus.Prepare HIV-1 by ordinary method
IIIB, titration also calculates viral TCID
50After the packing of virus stock solution, put-70 ℃ of preservations.Cell and virus frozen and recovery by ordinary method.
Four, HIV-1 infectious titration
HIV-1
IIIBCarry out titration by the said method improvement of Johnson&Byington (1990), be summarized as follows: the HIV-1 stock solution is done 4 times of dilutions on 96 orifice plates, 10 gradients, 6 repeating holes of every gradient are provided with control wells 6 holes simultaneously.Every hole adds C8166 cell 50 μ l, and every hole final volume is 200 μ l.37 ℃, 5%CO
2Cultivate.Added fresh RPMI-1640 perfect medium 100 μ l on the 3rd day, (whether Cytopathic Effect CPE), has the formation of synplasm (Syncytium) to confirm with every hole under inverted microscope, to observe HIV-1 inductive cytopathic effect in every hole on the 7th day; Press the TCID that the Reed&Muench method is calculated virus
50(50%Tissue Culture Infection Dose).
Five, to the toxicity test of C8166 cell
4 * 10
5/ ml C8166 cell suspension 100 μ l mix with different medicament solutions to be measured, establish 3 repeating holes.The not control wells of drug is set simultaneously, 37 ℃, 5%CO
2Cultivated 3 days, and adopted the MTT colourimetry to detect cytotoxicity.The ELx800 ELIASA is measured the OD value, and the mensuration wavelength is 595nm, and reference wavelength is 630nm.Calculate CC
50Value (50%CytotoxicConcentration), the drug level during promptly to 50% normal T lymphocyte series C8166 toxigenicity.
Six, to HIV-1
IIIBThe inhibition experiment of cytopathogenic effect (CPE)
With 8 * 10
5/ ml C8166 cell 50 μ l/ holes are inoculated on the 96 porocyte culture plates that contain 100 μ l/ hole gradient doubling dilution medicines, add the HIV-1 of 50 μ l then
IIIBThe dilution supernatant, the 1300TCID50/ hole.If 3 repeating holes.The not normal cell control wells of drug is set simultaneously.The positive medicine contrast of AZT.37 ℃, 5%CO
2Cultivated 3 days, plasmodial formation is counted in (100 *) under the inverted microscope.EC
50(50%Effective Concentration) forms 50% o'clock drug level for suppressing synplasm.
Seven, to HIV-1
IIIBInfect the protection experiment of MT-4 necrocytosis
On 96 porocyte culture plates, every kind of medicine is carried out 5 times of doubling dilutions, each extent of dilution is established 4 holes, totally 6 extent of dilution.Every hole 100 μ l.The not infection or the infected by HIV-1 of drug are set simultaneously
IIIBThe cell control group.AZT is as the positive control medicine.2 holes in each extent of dilution drip MT-4 cell suspension and HIV-1 respectively
IIIBEach 50 μ l (infection multiplicity is 0.3) of supernatant.2 holes drip MT-4 cell suspension 100 μ l in addition.Every hole final volume is 200 μ l.Put 37 ℃, cultivate in the 5%CO2 incubator.Infect the back and added 100 μ l fresh cultures on the 3rd day.Measured OD with the MTT method with the ELx800 ELIASA on the 5th day
595/630nmValue.Calculate the inhibiting rate of medicine cell growth and to HIV-1
IIIBThe protection ratio of cells infected.CC
50For to the toxigenous drug level of 50% host cell; EC
50For protecting 50% HIV-1
IIIBDrug level during the cells infected survival.
Eight, calculation formula
Draw dose response curve according to experimental result, calculate the 50% effective concentration (EC that compound suppresses virus by the Reed&Muench method
50), 50% cell growth inhibiting concentration (CC
50) and the active therapeutic index TI value of anti-HIV-1 (Therapeutic index) be: TI=CC
50/ EC
50
1, cell growth survival rate (%)=experimental port OD value/control wells OD value * 100
2, the cytopathogenic inhibiting rate of HIV-1 (%)=(1-experimental port synplasm number/control wells synplasm number) * 100
Nine, experimental result (seeing table 1).
For the first time the activity experiment result shows that medicine and virus hatches 2h earlier, the EC of Trigonostemon chinensis lactone A (1), Trigonostemon chinensis lactone B (2), Trigonostemon chinensis lactone C (3), Trigonostemon chinensis lactone D (4)
50Be respectively 1.62ng/ml, 6.46ng/ml,＜64ng/ml, 1.26ng/ml.
For the second time the screening active ingredients result shows that medicine and cell hatch 2h earlier, the EC of Trigonostemon chinensis lactone A (1), Trigonostemon chinensis lactone B (2), Trigonostemon chinensis lactone C (3), Trigonostemon chinensis lactone D (4)
50Be respectively 3.99ng/ml, 7.60ng/ml,＜64ng/ml, 2.37ng/ml.
Experimental result shows: above-claimed cpd has significant inhibition HIV and cell forms plasmodial effect, and has higher selectivity index.
The anti-HIV pharmacological tests of table 1 The compounds of this invention
Claims (5)
1. the Trigonostemon chinensis lactone A-D shown in the structural formula (I),
2. the inverase compsn contains compound and the pharmaceutically acceptable carrier shown in claim 1 formula (I) of treating significant quantity.
3. the preparation method of the said formula of claim 1 (I) compound gets the Meng Lun Trigonostemon chinensis, extracts three times with 95 % alcohol heating reflux; Each 4 hours, united extraction liquid, decompression recycling ethanol; Get medicinal extract, its water is pinched molten, filtration, use sherwood oil respectively; Ethyl acetate extraction three times; Get ethyl acetate extraction part, get Fr1, Fr2, Fr3, Fr4 four parts, wherein sherwood oil with sherwood oil-acetone gradient elution: 7: 3 wash-out parts of acetone Fr3 must contain the component of Trigonostemon chinensis lactone A-D with the SephadexLH-20 methanol-eluted fractions; This component recycle silicon plastic column chromatography chloroform: 8: 1 wash-outs of methyl alcohol, use the HPLC acetonitrile then: 65: 35 purifying of water get compound Trigonostemon chinensis lactone A, B, C and D.
4. the application of claim 1 formula (I) compound in the preparation anti-AIDS drug.
5. the application of the said pharmaceutical composition of claim 2 in the preparation anti-AIDS drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910218391A CN102101864B (en) | 2009-12-18 | 2009-12-18 | Trigonolactone A-D and medicinal composition, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910218391A CN102101864B (en) | 2009-12-18 | 2009-12-18 | Trigonolactone A-D and medicinal composition, preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102101864A CN102101864A (en) | 2011-06-22 |
| CN102101864B true CN102101864B (en) | 2012-09-05 |
Family
ID=44154953
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910218391A Active CN102101864B (en) | 2009-12-18 | 2009-12-18 | Trigonolactone A-D and medicinal composition, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102101864B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1368973A (en) * | 1999-07-01 | 2002-09-11 | 赞塔里斯股份公司 | Novel xanthone compounds, their preparation and use as medicament |
-
2009
- 2009-12-18 CN CN200910218391A patent/CN102101864B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1368973A (en) * | 1999-07-01 | 2002-09-11 | 赞塔里斯股份公司 | Novel xanthone compounds, their preparation and use as medicament |
Non-Patent Citations (1)
| Title |
|---|
| JP特开平8-92085A 1996.04.09 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102101864A (en) | 2011-06-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2017133468A1 (en) | Pulchinenoside and application as inhibitor of ev71 virus | |
| CN101732417B (en) | Preparation method and application of ion pair mixture of macleaya cordata total alkaloid | |
| CN105061456A (en) | Novel kaurene diterpenoid compound and preparation method and pharmaceutical application thereof | |
| CN102225937B (en) | Antineoplastic alkaloid compound, and its medicine composition, preparation method and application | |
| FI126047B (en) | Pharmaceutical compositions of anisomelic acid and their use | |
| CN106176716B (en) | The new application of daphane diterpene compound pimelotide C | |
| CN103880856B (en) | Bisabolane sesquiterpene derivant and pharmaceutical composition thereof and its application in pharmacy | |
| CN100497279C (en) | Pharmaceutical use of ent-eudesmane alcohol type sesquiterpene for inhibiting hepatitis virus | |
| CN105524063A (en) | Novel terpene indole alkaloid compound, and preparation method and medical application thereof | |
| CN102101864B (en) | Trigonolactone A-D and medicinal composition, preparation method and application thereof | |
| CN103012420B (en) | Lycorine derivative, veterinary drug taking lycorine derivative as an active constituent, and applications of lycorine derivative | |
| CN1118471C (en) | Medicine containing tan matter caesalpinia extract | |
| CN101239964A (en) | 15-oxospiramilactone and its medicament composition, preparation method and application | |
| CN103784427A (en) | Pharmaceutical composition containing sesquiterpene eudesmanolide and application of pharmaceutical composition in pharmaceuticals | |
| US10959974B2 (en) | Ingenol compounds and use thereof in anti-HIV latency treatment | |
| CN110818613B (en) | Carbazole compound, preparation method thereof and application thereof in anti-HIV (human immunodeficiency virus) medicines | |
| CN102584846B (en) | Curcumenol derivatives resisting influenza A(H1N1) virus | |
| CN103739575B (en) | 14-deoxidation-11,12-bis-andrographolide and pharmaceutical composition thereof and application | |
| CN102218047A (en) | Medicament for treating drug-resistant bacteria infection, and application of active ingredient thereof in pharmacy | |
| CN103739597B (en) | 14-deoxidation-14,15-bis-andrographolide and pharmaceutical composition thereof and purposes | |
| KR101988882B1 (en) | A composition comprising ginsenoside Rk1 or Rg5 for preventing or treating sepsis | |
| CN102796157B (en) | Caudatin derivative and medicinal composition and application thereof | |
| CN100338067C (en) | Anti-AIDS compound, its preparing method and pharmaceutical composition with anti-AIDS compound as active component and use thereof | |
| CN101829083B (en) | Application of substituted silybin ester in preparing medicament for treating virus hepatitis B | |
| CN101602787B (en) | Alisol A derivative, medicinal composition, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |