CN102276621B

CN102276621B - Compound separated from gamboge resin and derivative thereof and pharmaceutical composition comprising compound and derivative

Info

Publication number: CN102276621B
Application number: CN201010204256.6A
Authority: CN
Inventors: 李森彬
Original assignee: TAIWAN MORIMOTO BIOTECHNOLOGY DEVELOPMENT Co Ltd
Current assignee: TAIWAN MORIMOTO BIOTECHNOLOGY DEVELOPMENT Co Ltd
Priority date: 2010-06-11
Filing date: 2010-06-11
Publication date: 2015-01-07
Anticipated expiration: 2030-06-11
Also published as: CN102276621A

Abstract

The invention relates to a compound separated from gamboge resin and a derivative thereof and a pharmaceutical composition comprising the compound and the derivative, and discloses a novel composition with a chemical formula (I) shown in the specification, wherein each substituent is defined in specification and claims. The invention also discloses the compound, an acetone extraction product of the gamboge resin and application of a separated part from the acetone extraction product to preparation of the pharmaceutical composition.

Description

Be separated the compound from Gamboge and its derivative, and include the pharmaceutical composition of these compounds and derivative

Technical field

The present invention relates to one, there is the separate part of the acetone extract product (acetone-extracted product) being obtained from Gamboge (gamboge resin) about 5 kinds, and by 18 kinds of compounds being further purified out and its derivative from this acetone extract product, particularly relate to these separate parts a kind of and these compounds are proved the activity with Tumor suppression/growth of cancer cells.This acetone extract product and these separate parts are also proved the activity with pain relieving and anti-inflammation.Also have about this acetone extract product, these separate parts and these compounds and its derivative.

background technology

Gamboge (gamboge resin) is the colloid resin (gum-resin) secreted by good fortune Ochnaceae (Guttiferae) garcinia plant (plant of Garcinia sp.), namely it be used as the source of vegetable dyes and pigment since ancient times, is also that some areas are as the Folk medicine (folk-medicine) of the state such as India, Thailand.

(Classification system is gamboge: Garcinia morella Desv; The Chinese phonetic alphabet is TENGHUANG; English by name gamboge) be aiphyllium, be grown on torrid areas, be distributed in India person and be mainly Garcinia morelll Desv, and be distributed in Thailand person and be mainly G.hanburyi Hook f..Before blooming, the skin zone of stem cuts open as spirrillum and collects the resin of outflow by liftoff about 2 meters of, namely becomes the Gamboge (gamboge resin) of solid state after being heated evaporate to dryness (heat-drying).According to the record of traditional Chinese medicine (Traditional Chinese Medicine, TCM) ancient books and records, gamboge (gamboge) has effect of detumescence, change poison, hemostasis, desinsection.From 1934 so far, there is the research of ingredient in many reported in literature Gamboges, and known chemical compound lot of can isolating from the extract of Gamboge at present, comprising: morellin (morellin), morellic acid (morellic acid), gambogic acid (gambogic acid), gamboge alcohol (morellinol), isomorellin (isomorellin), isomorellic acid (isomorellic acid), Isogambogic acid (isogambogic acid), isomorellinol (isomorellinol), neogambogic acid (neogambogic acid), desoxymorellin (desoxymorellin), dihydro isomorellin (dihydroisomorellin), gambogenic acid (gambogenic acid), desoxygambogenin (desoxygambogenin), gambogellic acid (gambogellic acid), epigambogic acid (epigambogic acid), epiisogambogic acid (epiisogambogic acid), isogambogenic acid (isogambogenic acid) and 30-hydroxygambogic acid (30-hydroxygambogic acid) etc.

Once reported in literature was had, Gamboge contains some composition for human cervical cancer cell HeLa (human cervical cancer cells HeLa), human nasopharynx's cancer cells KB (humannasopharyngeal cancer cells KB), human leukaemia cell K562 (human leukemiacells K562) and anti-many ropes are as pungent K562 cell strain (doxorubicin-resistant K562cell lines) etc. having cytotoxicity (cytotoxic activity) (J.Asano et al. (1996), Phytochemistry, 41:815-820, L.J.Lin et al. (1993), MagneticResonance in Chemistry, 31:340-347, Q.B.Han et al. (2006), PlantaMed., 72:281-284, Q.B.Han et al. (2006), Chem.Pharm.Bull., 54:265-267, Q.B.Han et al. (2006), Chemistry & Biodiversity, 3:101-105).

US 6,462,041 B1 discloses with the gambogic acid of following represented by formula I (gambogicacid) and its analogue and derivative:

[wherein dotted line is singly-bound, double bond or cycloalkyl groups (epoxy groups); And X, Y and R1 to R3 as in this case define]

But the overall disclosure of US 6,462,041 B1 does not illustrate compound with Formula I containing 32,33-cycloalkyl groups and preparation method thereof.In addition, in this Patent Case, disclose the above-mentioned compound with Formula I is the inducer that activator and the property planned of half Guang-aspartyl protease (caspases) carves die (apoptosis).

In authorizing the TW I282280 of the people such as Chen Qiuming (corresponding to US 7, 138, 428B2 and CN100413868C) in disclose one be obtained from Gamboge acetone extract product TSB-14 (acetone-extracted product TSB-14) and from this acetone extract product TSB-14 by the compound be further purified out, comprise betula camphor (betulin), Betulinic Acid (betulinicacid), morellic acid, isomorellic acid, gambogic acid, Isogambogic acid, isomorellinol, desoxymorellin and one is named as the new compound of formoxanthone A (formoxanthone A).This acetone extract product and these be proved the effectiveness that there is Tumor suppression/cancer cells [such as liver cancer cell (HepG2), lung carcinoma cell (A549), breast cancer cell (MCF-7), colorectal cancer cells (HT-29), leukemia cell (HL-60) and lymphoma cell (U937) etc.] and grow by the compound be further purified.

US 2007/0093456A1 discloses a kind of Gamboge acid derivative, and it is named as 37,38-dihydroxyl-gambogic acid methyl ester (methyl 37,38-dihydroxy-gambogate) and has chemical structural formula as shown below.Find via experiment, 37,38-dihydroxyl-gambogic acid methyl ester can carve the inducer of dying as the activator of half Guang-aspartyl protease and the property planned.

(37,38-dihydroxyl-gambogic acid methyl ester)

At S.J.Tao et al. (2009), Journal of Natural Products, in 72:117-124, the people such as S.J.Tao isolate 12 kinds of novel yellow pigment (xanthones) [such as oxygen base gambogic acid (oxygambogic acid) from the resin of gamboge (G.hanburyi), 8, 8a-dihydro morellic acid methyl ester (methyl 8, 8a-dihydromorellate) and 7-methoxyl group gambogellic acid (7-methoxygambogellic acid) etc.] and the natural product [namely 8 of a pair novelty, 8a-dihydro-8-hydroxyl gambogic acid (8, 8a-dihydro-8-hydroxygambogic acid) and its isomer (isomer)], wherein three-dimensional arrangement (stereostructure) method of there is no of oxygen base gambogic acid is determined, the configuration of C15/C16 double bond (configuration) is caused to be judged out because of H-15 and the H-16 signal overlap in 1H-NMR spectrum.In addition, the people such as S.J.Tao find via pharmacological evaluation, and outside 8,8a-dihydro morellic acid methyl ester, other 13 kinds of compounds all have the effectiveness suppressing HeLa growth of tumour cell.

Except the activity of Tumor suppression/growth of cancer cells, the extract of Gamboge is also proved (oxygen base gambogic acid)

There is other biological activity.Such as, at A.Panthong et al. (2007), Journalof Ethnopharmacology, in 111:335-340, the people such as A.Panthong disclose the acetic acid ethyl ester extract (ethyl acetate extract) (it is named as GH5763) that one is derived from the resin of gamboge (G.hanburyi Hook f.), and find via experimentation on animals, this acetic acid ethyl ester extract GH5763 has the activity of anti-inflammation (anti-inflammatory), pain relieving (analgesic) and antipyretic (antipyretic).

Although as mentioned above, for the Pharmaceutical Chemist in pharmacy industry and producer, still have needs go development easily to be prepared and there is pain relieving, the compounds of anti-inflammation and antitumour activity (analgesic, anti-inflammatory and anticancer activities) or extract.

After deliberation, applicant finds that the acetone extract product TSB-14 of the Gamboge disclosed in TW I282280 is except having the activity of Tumor suppression/growth of cancer cells, also has the activity of pain relieving and anti-inflammation.In addition, applicant isolates 5 separate parts (fractions) further and is purified into 18 kinds of compounds from this acetone extract product TSB-14, find after deliberation, these separate parts have the activity of Tumor suppression/growth of cancer cells, pain relieving and anti-inflammation, and these compounds also have the activity of Tumor suppression/growth of cancer cells.

As can be seen here, above-mentioned existing Gamboge is at method, product and use, and obviously still has inconvenience and defect, and is urgently further improved.In order to solve above-mentioned Problems existing, relevant manufactures there's no one who doesn't or isn't seeks solution painstakingly, but have no applicable design for a long time to be completed by development, and general method and product do not have appropriate method to solve the problem, this is obviously the anxious problem for solving of relevant dealer always.Therefore how to found a kind of separation newly from the compound of Gamboge and its derivative, and include the pharmaceutical composition of these compounds and derivative, real one of the current important research and development problem that belongs to also becomes the target need improved current industry pole.

Summary of the invention

The object of the invention is to, the compound of a kind of separation newly from Gamboge and its derivative are provided, and including the pharmaceutical composition of these compounds and derivative, technical problem to be solved makes it provide one to have the compound of following chemical formula (I);

There is provided a kind of for suppressing the pharmaceutical composition of the growth of a lesion/cancer cell;

A kind of pharmaceutical composition being used for the treatment of a lesion/cancer disease is provided;

A kind of pharmaceutical composition with analgesic activity is provided;

A kind of pharmaceutical composition with anti-inflammation activity is provided;

Applicant finds that the acetone extract product TSB-14 of the Gamboge disclosed in TW I282280 is except having the activity of Tumor suppression/growth of cancer cells, also has the activity of pain relieving and anti-inflammation.In addition, applicant isolates 5 separate parts (fractions) further and is purified into 18 kinds of compounds from this acetone extract product TSB-14, find after deliberation, these separate parts have the activity of Tumor suppression/growth of cancer cells, pain relieving and anti-inflammation, and these compounds also have the activity of Tumor suppression/growth of cancer cells.

The object of the invention to solve the technical problems realizes by the following technical solutions.The one proposed according to the present invention has the compound of following chemical formula (I):

Wherein

R1 and R2 forms one together and is selected from one the part represented by the chemical formula in following formed group:

R3 is selected from by following formed group: 3-methyl-2-butene base, 1-hydroxyl-2,3-epoxy group(ing)-3-methyl butyl, 2,3-epoxy group(ing)-3-methyl butyl, 2,3-dihydroxyl-3-methyl butyls, 3-hydroxy-3-methyl butyl, 3-hydroxy-3-methyl-1E-butenyl, 3-hydroxy-3-methyl-1Z-butenyl, 2-hydroxy-3-methyl-3-butenyl and formylethyl; And

R4 is selected from by following formed group: 2-carboxyl-2Z-butenyl, 2-carboxyl-2E-butenyl, 2-carboxyl-3Z-butenyl and 2-carboxyl-3E-butenyl,

But with good conditionsi, following compounds is excluded:

The object of the invention to solve the technical problems also can be applied to the following technical measures to achieve further.

Aforesaid compound, wherein said R3 is selected from by following formed group: 3-methyl-2-butene base, 1-hydroxyl-2,3-epoxy group(ing)-3-methyl butyl, 2,3-epoxy group(ing)-3-methyl butyls and 2,3-dihydroxyl-3-methyl butyl; And

R4 is selected from by following formed group: 2-carboxyl-2Z-butenyl, 2-carboxyl-2E-butenyl and 2-carboxyl-3E-butenyl.

Aforesaid compound, wherein said when R 3 be 3-methyl-2-butene base and R4 is 2-carboxyl-2Z-butenyl, R1 and R2 forms one together and is selected from one the part represented by the chemical formula in following formed group:

When R3 is 1-hydroxyl-2,3-epoxy group(ing)-3-methyl butyl, 2, when 3-epoxy group(ing)-3-methyl butyl or 2,3-dihydroxyl-3-methyl butyl and R4 are 2-carboxyl-2Z-butenyls, R1 and R2 forms one together and is selected from one the part represented by the chemical formula in following formed group:

When R4 is 2-carboxyl-2E-butenyl, R3 is 3-methyl-2-butene base, and R1 and R2 forms a part represented with following chemical formula together:

; And

When R4 is 2-carboxyl-3E-butenyl, R3 is 3-methyl-2-butene base, and R1 and R2 forms one together and is selected from one the part represented by the chemical formula in following formed group:

The object of the invention to solve the technical problems also realizes by the following technical solutions.It is a kind of for suppressing the pharmaceutical composition of the growth of a lesion/cancer cell that foundation the present invention proposes, wherein

Pharmaceutical compositions include following any one: a compound as claimed in claim 1 with chemical formula (I); And one is selected from by the separate part being obtained from an acetone extract product of Gamboge in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E, wherein separate part TSB-14A be in fact by formoxanthone A,

Separate part TSB-14B is made up of isomorellic acid, morellic acid and isomorellinol;

Separate part TSB-14C be in fact by 30-hydroxygambogic acid, 30-hydroxyepigambogic acid, neogambogic acid,

Separate part TSB-14D be in fact by isomorellin, isogambogenic acid, gambogenic acid and

formed; And

Separate part TSB-14E be in fact by gambogellic acid, Isogambogic acid, epiisogambogic acid, gambogic acid, epigambogic acid, desoxymorellin, desoxygambogenin,

institute

The object of the invention to solve the technical problems also adopts following technical measures to realize in addition.

Aforesaid pharmaceutical composition, wherein said lesion/cancer cell is selected from by following formed group: myelogenous leukemia cells, human hepatocytes cancer cells, Human Lung Cancer cell and human tissue cells's property lymphoma cell before mankind mastopathy cell, human colon's adenocarcinoma cell, the mankind.

The object of the invention to solve the technical problems also realizes in addition by the following technical solutions.According to a kind of pharmaceutical composition being used for the treatment of a lesion/cancer disease that the present invention proposes, wherein pharmaceutical compositions includes one and is selected from by the separate part being obtained from an acetone extract product of Gamboge in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E

Wherein separate part TSB-114A be in fact by formoxanthone A,

formed; And separate part TSB-14E be in fact by gambogellic acid, Isogambogic acid, epiisogambogic acid, gambogic acid, epigambogic acid, desoxymorellin, desoxygambogenin,

The wherein said lesion/cancer cell of aforesaid pharmaceutical composition is selected from by following formed group: colorectal carcinoma, cancer of the stomach, nonsmall-cell lung cancer, the cancer of the brain, thyroid carcinoma, nasopharyngeal carcinoma, chronic lymphocytic leukemia, T-cell acute lymphoid blast cell leukemia, lung cancer, colorectal carcinoma, breast cancer, leukemia, liver cancer, ovarian cancer, renal cancer, cancer of pancreas and carcinoma of endometrium.

The object of the invention to solve the technical problems also realizes in addition by the following technical solutions.According to a kind of pharmaceutical composition being used for the treatment of a lesion/cancer disease that the present invention proposes, wherein pharmaceutical composition includes an acetone extract product of Gamboge, and wherein this lesion/cancer disease is selected from by following formed group: colorectal carcinoma, cancer of the stomach, nonsmall-cell lung cancer, the cancer of the brain, thyroid carcinoma, nasopharyngeal carcinoma, chronic lymphocytic leukemia, T-cell acute lymphoid blast cell leukemia, leukemia, ovarian cancer, renal cancer, cancer of pancreas and carcinoma of endometrium.

The object of the invention to solve the technical problems also realizes in addition by the following technical solutions.According to the present invention propose a kind of pharmaceutical composition with analgesic activity, wherein pharmaceutical compositions include following any one: an acetone extract product of Gamboge; And one is selected from by the separate part being obtained from an acetone extract product of Gamboge in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E, wherein separate part TSB-14A be in fact by formoxanthone A,

formed; And

The object of the invention to solve the technical problems realizes in addition more by the following technical solutions.According to the present invention propose a kind of pharmaceutical composition with anti-inflammation activity, wherein pharmaceutical compositions include following any one: an acetone extract product of Gamboge; And one is selected from by the separate part being obtained from an acetone extract product of Gamboge in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E, wherein separate part TSB-14A be in fact by formoxanthone A,

formed;

formed; And

formed.

Aforesaid pharmaceutical composition, wherein said pharmaceutical compositions includes a pharmaceutically acceptable supporting agent further.

Aforesaid pharmaceutical composition, wherein said pharmaceutically acceptable supporting agent comprises one or more following reagent: solvent, emulsifying agent, suspension agent, decomposition agent, binding agent, vehicle, tranquilizer, sequestrant, thinner, jelling agent, sanitas, lubricant, absorption delay agent, fluidizer, weighting agent, disintegrating agent, surfactants, thickening material and liposome.

The present invention compared with prior art has obvious advantage and beneficial effect.From above technical scheme, main technical content of the present invention is as follows: by technique scheme, and the present invention is separated from the compound of Gamboge and its derivative, and includes the pharmaceutical composition of these compounds and derivative

One is provided to have the compound of following chemical formula (I);

A kind of pharmaceutical composition with analgesic activity is provided;

Above-mentioned explanation is only the general introduction of technical solution of the present invention, in order to technique means of the present invention can be better understood, and can be implemented according to the content of specification sheets, and can become apparent to allow above and other objects of the present invention, feature and advantage, below especially exemplified by preferred embodiment, and coordinate accompanying drawing, be described in detail as follows.

Accompanying drawing explanation

The elution figure that the acetone extract product TSB-14 that Fig. 1 shows the method using the embodiment 1 according to TW I282280 to disclose and the Gamboge be made obtains to carry out AG RP-HPLC, its medium wave peak 1 to crest 35 represents 35 major ingredient occurred between the residence time of the 0th to the 80th minute;

The acetone extract product TSB-14 that Fig. 2 shows the method using the embodiment 1 according to TW I282280 to disclose and the Gamboge be made carries out the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 1 to crest 35 corresponds respectively to crest 1 to the crest 35 of Fig. 1;

Fig. 3 shows to use and carries out according to separate part 1 obtained by embodiments of the invention 2 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 1 to crest 12 corresponds respectively to crest 1 to the crest 12 of Fig. 2;

Fig. 4 shows to use and carries out according to separate part 2 obtained by embodiments of the invention 2 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 13 to crest 24 corresponds respectively to crest 13 to the crest 24 of Fig. 2;

Fig. 5 shows to use and carries out according to separate part 3 obtained by embodiments of the invention 2 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 25 to crest 35 corresponds respectively to crest 25 to the crest 35 of Fig. 2;

The acetone extract product TSB-14 that Fig. 6 shows the method using the embodiment 1 according to TW I282280 to disclose and the Gamboge be made carries out the elution figure that half preparation scale RP-HPLC obtains;

Fig. 7 shows to use and carries out according to separate part TSB-14A obtained by embodiments of the invention 4 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 1 to crest 12 corresponds respectively to crest 1 to the crest 12 of Fig. 1;

Fig. 8 shows to use and carries out according to separate part TSB-14B obtained by embodiments of the invention 4 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 13 to crest 15 corresponds respectively to crest 13 to the crest 15 of Fig. 1;

Fig. 9 shows to use and carries out according to separate part TSB-14C obtained by embodiments of the invention 4 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 16 to crest 20 corresponds respectively to crest 16 to the crest 20 of Fig. 1;

Figure 10 shows to use and carries out according to separate part TSB-14D obtained by embodiments of the invention 4 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 21 to crest 24 corresponds respectively to crest 21 to the crest 24 of Fig. 1; And

Figure 11 shows to use and carries out according to separate part TSB-14E obtained by embodiments of the invention 4 the elution figure that half preparation scale RP-HPLC obtains, and its medium wave peak 25 to crest 35 corresponds respectively to crest 25 to the crest 35 of Fig. 1.

Embodiment

For further setting forth the present invention for the technique means reaching predetermined goal of the invention and take and effect, below in conjunction with accompanying drawing and preferred embodiment, to the separation proposed according to the present invention from the compound of Gamboge and its derivative, and include its embodiment of pharmaceutical composition of these compounds and derivative, method steps, feature and effect thereof, be described in detail as follows.As used in this article, term " derivative " means the compound once chemically modified, and wherein this chemically modified occurs in functional group's group (functional group) or the aromatic ring (aromatic ring) of this compound.

Applicant at previous Taiwan Patent case TW I282280 (corresponding to US 7, 138, 428B2 and CN 100413868 C) in disclosed gambogic acid acetone extract product TSB-14 include 9 kinds of compounds, comprise: formoxanthone A (formoxanthone A), betula camphor (betulin), Betulinic Acid (betulinic acid), morellic acid (morellic acid), isomorellic acid (isomorellicacid), gambogic acid (gambogic acid), Isogambogic acid (isogambogic acid), isomorellinol (isomorellinol) and desoxymorellin (desoxymorellin).For confirming whether this acetone extract product TSB-14 has other activeconstituents further, applicant uses AG (analytical) reverse phase high performance energy liquid chromatography (LC) (reversed phase high performanceliquid chromatography, RP-HPLC) and half preparation scale (semipreparative) RP-HPLC to carry out the isolation and purification of composition.

This acetone extract product TSB-14 first carries out AG RP-HPLC with AG RP-C8 tubing string [Luna 3 μ C8 (2)], and obtain the AG RP-HPLC elution figure (as shown in Fig. 1 of embodiment 1 below) of this acetone extract product TSB-14, wherein this AG RP-HPLC elution graphics package contains 35 main elution crests (being denoted as crest 1 to crest 35 respectively).

This acetone extract product TSB-14 then carries out half preparation scale RP-HPLC with half preparation scale RP-Cl2 tubing string (Synergi 4 μ Cl2), and obtains the half preparation scale RP-HPLC elution figure (as shown in Fig. 2 of embodiment 2 below) of this acetone extract product TSB-14.Can know after the AG RP-HPLC elution figure of this half preparation scale RP-HPLC elution figure and above-mentioned acetone extract product TSB-14 is done a comparison and find out the position of crest 1 to crest 35 in half preparation scale RP-HPLC elution figure.These 35 crests are distinguished into 3 sections (sections) (namely, section 1 to section 3) by applicant, and wherein section 1 is containing crest 1 to crest 12, and the residence time is the 0th to the 42nd minute; Section 2 is containing crest 13 to crest 24, and the residence time is the 42nd to the 135th minute; And section 3 is containing crest 25 to crest 35, the residence time is the 135th to the 280th minute.

Afterwards, this acetone extract product TSB-14 is carried out half preparation scale RP-HPLC, and when elution, collect the washings (eluate) corresponding to each section respectively according to section 1 to the section 3 of the half preparation scale RP-HPLC elution figure of above-mentioned acetone extract product TSB-14.Each washings carries out distribution with H2O with ethyl acetate (ethyl acetate) respectively and is separated (partitioning), then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener (vacuum rotatory evaporator), and obtain separate part 1, separate part 2 and separate part 3.Then, these separate part 1 to separate parts 3 are carried out half preparation scale RP-HPLC respectively, and obtain the half preparation scale RP-HPLC elution figure (as shown in Fig. 3 to Fig. 5 of embodiment 2 below) of separate part 1 to separate part 3, wherein half preparation scale RP-HPLC elution figure of separate part 1 to separate part 3 is respectively containing crest 1 to crest 12, crest 13 to crest 24 and crest 25 to crest 35.

This separate part 1 is carried out half preparation scale RP-HPLC, and when elution, collect according to half preparation scale RP-HPLC elution figure of above-mentioned separate part 1 (namely below embodiment 2 Fig. 3) washings 1 to the washings 12 corresponding respectively to crest 1 to crest 12.Washings 1 to washings 12 carries out distribution with H2O with ethyl acetate respectively and is separated, and then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener, and obtain crude product Gh-3261, Gh-3271, Gh-3272, Gh-3311, Gh-3332, Gh-1036, Gh-3291, Gh-631, Gh-1052, Gh-3351, Gh-3353 and Gh-3352.These 12 kinds of crude products are carried out AG RP-HPLC respectively, and obtains the AG RP-HPLC elution figure of each crude product.If the AG RP-HPLC elution figure of the crude product obtained does not demonstrate single crest, AG RP-HPLC elution figure remaining crude product repeated half above-mentioned preparation scale RP-HPLC to carry out further purifying, until can demonstrate single crest (D item and table 1 with reference to embodiment 2 below).

This separate part 2 is carried out half preparation scale RP-HPLC, and when elution, collect according to half preparation scale RP-HPLC elution figure of above-mentioned separate part 2 (namely below embodiment 2 Fig. 4) washings 13 to the washings 24 corresponding respectively to crest 13 to crest 24.Washings 13 to the washings 24 obtained connects and carries out distributions isolation and purification according to aforesaid method, and obtains product Gh-47, Gh-4602, Gh-4601, Gh-1601-A, Gh-1050, Gh-1602, Gh-1631, Gh-2641-1, Gh-2501, Gh-2642, Gh-2507 and Gh-2505 (reference is the E item of embodiment 2 and table 2 below).

This separate part 3 is carried out half preparation scale RP-HPLC, and when elution, collect according to the AG RP-HPLC elution figure of above-mentioned separate part 3 (namely below embodiment 2 Fig. 5) washings 25 to the washings 35 corresponding respectively to crest 25 to crest 35.Washings 25 to the washings 35 obtained connects and carries out distributions isolation and purification according to aforesaid method, and obtains product Gh-2508, Gh-2603-1, Gh-2603-2, Gh-1641, Gh-1642, Gh-2605, Gh-2606, Gh-2607-B, Gh-2607-1A, Gh-2301 and Gh-4301 (reference is the F item of embodiment 2 and table 3 below).

The product of above-mentioned 35 kinds of purifying self-separation part 1 to separate parts 3 is carried out physics and chemistry analysis, comprise: fusing point, NMR (Nuclear Magnetic Resonance) spectrum (nuclear magnetic resonancespectroscopy) (such as 1H-NMR and 13C-NMR spectrum) and mass spectrum (massspectrometry) [such as electron bombardment mass spectrum (electron impact mass spectrometry, EIMS), high parsing electron bombardment mass spectrum (high-resolution electron impact massspectrometry, HREIMS), fast atom bombardment mass spectrum (fast atom bombardment massspectrometry, and high resolve fast atom bombardment mass spectrum (high-resolutionfast atom bombardment mass spectrometry FABMS), HRFABMS)] etc. (with reference to embodiment 3 below).

It is a known compound with following shown chemical structural formula that product Gh-631 is identified after Identification of chemical structure, namely formoxanthone A (formoxanthone A):

gh-631 (formoxanthone A)

It is 2 kinds of known morellic acid stereoisomerses that product Gh-4602 and Gh-47 is identified after Identification of chemical structure, namely morellic acid (morellic acid) and isomorellic acid (isomorellicacid), they have following shown chemical structural formula respectively:

It is a kind of known isomorellic acid derivative, namely isomorellinol (isomorellinol) that product Gh-4601 is identified after Identification of chemical structure, and it has following shown chemical structural formula:

gh-4601 (isomorellinol)

It is the C-2 epimer (C-2epimers) of 2 kinds of known 30-hydroxygambogic acids that product Gh-1601-A and Gh-1602 is identified after Identification of chemical structure, namely 30-hydroxygambogic acid (30-hydroxygambogic acid) and 30-hydroxyepigambogic acid (30-hydroxyepigambogic acid), they have following shown chemical structural formula respectively:

gh-1601-A (30-hydroxygambogic acid)

gh-1602 (30-hydroxyepigambogic acid)

It is a known compound with following shown chemical structural formula that product Gh-2641-1 is identified after Identification of chemical structure, namely neogambogic acid (neogambogic acid):

gh-2641-1 (neogambogic acid)

It is a known compound with following shown chemical structural formula that product Gh-2501 is identified after Identification of chemical structure, namely isomorellin (isomorellin):

gh-2501 (isomorellin)

It is 2 kinds of known gambogenic acid stereoisomerses that product Gh-2505 and Gh-2642 is identified after Identification of chemical structure, namely gambogenic acid (gambogenic acid) and isogambogenic acid (isogambogenic acid), they have following shown chemical structural formula respectively:

gh-2505 (gambogenic acid)

gh-2642 (isogambogenic acid)

It is a known compound with following shown chemical structural formula that product Gh-2603-2 is identified after Identification of chemical structure, namely gambogellic acid (gambogellic acid):

gh-2603-2 (gambogellic acid)

It is the C-2 epimer of 2 kinds of known Isogambogic acids that product Gh-1641 and Gh-1642 is identified after Identification of chemical structure, namely Isogambogic acid (isogambogic acid) and epiisogambogic acid (epiisogambogic acid), they have following shown chemical structural formula respectively:

It is the C-2 epimer of 2 kinds of known gambogic acids that product Gh-2605 and Gh-2606 is identified after Identification of chemical structure, namely gambogic acid (gambogic acid) and epigambogic acid (epigambogic acid), they have following shown chemical structural formula respectively:

It is a known compound with following shown chemical structural formula that product Gh-2301 is identified after Identification of chemical structure, namely desoxymorellin (desoxymorellin):

gh-2301 (desoxymorellin)

It is a known compound with following shown chemical structural formula that product Gh-4301 is identified after Identification of chemical structure, namely desoxygambogenin (desoxygambogenin):

gh-4301 (desoxygambogenin)

Product Gh-3352 and Gh-3351 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as formoxanthone E (formoxanthone E) and epiformoxanthone E (epiformoxanthone E) respectively, and have following shown chemical structural formula respectively:

Product Gh-1052 and Gh-1036 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as formoxanthone F (formoxanthone F) and epiformoxanthone F (epiformoxanthone F) respectively, and have following shown chemical structural formula respectively:

Product Gh-3353 and Gh-3311 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as formoxanthone G (formoxanthone G) and piformoxanthone G (epiformoxanthone G) respectively, and have following shown chemical structural formula respectively:

Product Gh-3261 and Gh-3271 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as formoxanthone J (formoxanthone J) and epiformoxanthone J (epiformoxanthone J) respectively, and have following shown chemical structural formula respectively:

Product Gh-3272 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as formoxanthone H (formoxanthone H), and has following shown chemical structural formula:

gh-3272 (formoxanthone H)

Product Gh-3332 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as isoformoxanthone I (isoformoxanthone I), and has following shown chemical structural formula:

gh-3332 (isoformoxanthone I)

Product Gh-3291 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as formoxanthone D (formoxanthone D), and has following shown chemical structural formula:

gh-3291 (formoxanthone D)

Product Gh-1631 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as formoxanthone C (formoxanthone C), and has following shown chemical structural formula:

gh-1631 (formoxanthone C)

Product Gh-1050 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as 3 Alpha-hydroxy gambogellic acids (3 α-hydroxygambogellic acid), and has following shown chemical structural formula:

Gh-1050 (3 Alpha-hydroxy gambogellic acid)

Product Gh-2603-1 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be a compounds not seen that document recorded, it is named as epigambogellic acid (epigambogellic acid), and has following shown chemical structural formula:

Gh-2603-1 (epigambogellic acid)

Product Gh-2508 and Gh-2507 via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as β-gambogellic acid (β-gambogellic acid) and β-epigambogellic acid (β-epigambogellic acid) respectively, and have following shown chemical structural formula respectively:

Product Gh-2607-B and Gh-2607-1A via Identification of chemical structure and with the SPECTRAL DATA comparison of known compound after to be identified be 2 kinds of compounds and C-2 epimer not seen that document recorded, they are named as formoxanthone B (formoxanthone B) and epiformoxanthone B (epiformoxanthone B) respectively, and have following shown chemical structural formula respectively:

The chemical structure of the above-mentioned 18 kinds of compounds of applicant's comparison finds, these compounds all have identical skeleton structure (skeleton structure).Therefore, the invention provides the compound that one has following chemical formula (I):

Wherein:

But with good conditionsi, following compounds is excluded:

Preferably, R3 representative: 3-methyl-2-butene base, 1-hydroxyl-2,3-epoxy group(ing)-3-methyl butyl, 2,3-epoxy group(ing)-3-methyl butyls and 2,3-dihydroxyl-3-methyl butyl.

Preferably, R4 representative: 2-carboxyl-2Z-butenyl, 2-carboxyl-2E-butenyl and 2-carboxyl-3E-butenyl.

Include, but are not limited to according to the typical example with the compound of chemical formula (I) of the present invention: epigambogellic acid, formoxanthone B, epiformoxanthone B, β-gambogellic acid, β-epigambogellic acid, formoxanthone C, 3 Alpha-hydroxy gambogellic acids, formoxanthone D, formoxanthone E, epiformoxanthone E, formoxanthone F, epiformoxanthone F, formoxanthone G, piformoxanthone G, formoxanthone H, isoformoxanthone I, formoxanthone J and epiformoxanthone J.

Can adopt according to the compound with chemical formula (I) of the present invention and have the knack of this those skilled in the art and know in detail and usual chemical synthesis process and being produced.

In addition, applicant uses different operational conditions to carry out the half preparation scale RP-HPLC of this acetone extract product TSB-14, and obtains the another kind half preparation scale RP-HPLC elution figure (as shown in Fig. 6 of embodiment 4 below) of this acetone extract product TSB-14.This half preparation scale RP-HPLC elution figure is divided into 5 sections (namely, section A is to section E) by applicant, and wherein the residence time of section A is the 0th to the 13.7th minute; The residence time of section B is the 13.7th to the 20.4th minute; The residence time of section C is the 20.4th to the 26.1st minute; The residence time of section D is the 26.1st to the 34.9th minute; And the residence time of section E is the 34.9th to the 53.8th minute.

Afterwards, this acetone extract product TSB-14 is carried out half preparation scale RP-HPLC, and when gradient elution, collect the washings corresponding to each section respectively according to the section A to section E of the half preparation scale RP-HPLC elution figure of above-mentioned acetone extract product TSB-14.Each washings carries out distribution with H2O with ethyl acetate respectively and is separated, and then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener, and obtain roughing out part TSB-14A, TSB-14B, TSB-14C, TSB-14D and TSB-14E.Then, these roughing out part TSB-14A to separate part TSB-14E is repeated half preparation scale RP-HPLC to carry out further purifying, until half preparation scale RP-HPLC elution figure of each separate part all presents consistent situation (the C item see embodiment 4 below).These purified separate part TSB-14A to separate part TSB-14E are carried out half preparation scale RP-HPLC respectively, and obtains the half preparation scale RP-HPLC elution figure (as shown in Fig. 7 to Figure 11 of embodiment 4 below) of purified separate part TSB-14A to separate part TSB-14E.Can know after the half preparation scale RP-HPLC elution figure of purified separate part TSB-14A to separate part TSB-14E is done a comparison with the AG RP-HPLC elution figure of above-mentioned acetone extract product TSB-14 (as shown in Fig. 1 of embodiment 1 below) respectively and find out, the half preparation scale RP-HPLC elution figure of purified separate part TSB-14A to separate part TSB-14E is respectively containing crest 1 to crest 12, crest 13 to crest 15, crest 16 to crest 20, crest 21 to crest 24 and crest 25 to crest 35.This represents that purified separate part TSB-14A contains product Gh-3261, Gh-3271, Gh-3272, Gh-3311, Gh-3332, Gh-1036, Gh-3291, Gh-631, Gh-1052, Gh-3351, Gh-3353 and Gh-3352; Purified separate part TSB-14B contains product Gh-47, Gh-4602 and Gh-4601; Purified separate part TSB-14C contains product Gh-1601-A, Gh-1050, Gh-1602, Gh-1631 and Gh-2641-1; Purified separate part TSB-14D contains product Gh-2501, Gh-2642, Gh-2507 and Gh-2505; And purified separate part TSB-14E contains product Gh-2508, Gh-2603-1, Gh-2603-2, Gh-1641, Gh-1642, Gh-2605, Gh-2606, Gh-2607-B, Gh-2607-1A, Gh-2301 and Gh-4301.

Applicant further by vitro anticancer test (in vitro anti-cancer test) study 5 kinds of obtaining according to the present invention be derived from the acetone extract product of Gamboge separate part TSB-14A to separate part TSB-14E and by separate part 1 to separate part 3 by the antitumour activity of 18 kinds of compounds be further purified out.Be found to via experimental result: these separate parts all have the activity that Tumor suppression/cancer cells (before such as people Class breast cancer cell, human colon's adenocarcinoma cell, the mankind myelogenous leukemia cells, human hepatocytes cancer cells, Human Lung Cancer cell and human tissue cells's property lymphoma cell) grows.Especially, the effectiveness of these separate parts TSB-14A to separate part TSB-14E on Tumor suppression/growth of cancer cells is all good than the acetone extract product TSB-14 institute tool person of Gamboge haply.In addition, these 18 kinds of compounds also have the activity that Tumor suppression/cancer cells (before such as people Class breast cancer cell, human colon's adenocarcinoma cell, the mankind myelogenous leukemia cells, human hepatocytes cancer cells, Human Lung Cancer cell and human tissue cells's property lymphoma cell) grows, and applicant is inference accordingly: have the activity that a compound being similar to the skeleton structure of these 18 kinds of compound institute tool persons also has Tumor suppression/growth of cancer cells.

Based on above-mentioned, the compound that the separate part TSB-14A to separate part TSB-14E and of the acetone extract product obtained according to the present invention has chemical formula (I) is as above expected the potentiality with the growth that can be used for suppression one lesion/cancer cell.

Therefore, the invention provides a kind of for suppressing the pharmaceutical composition of the growth of a lesion/cancer cell, its include following any one:

One compound as above with chemical formula (I); And

One is selected from by the separate part being obtained from an acetone extract product of Gamboge in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E,

Wherein separate part TSB-14A be in fact by formoxanthone A,

Separate part TSB-14D be in fact by isomorellin, isogambogenic acid, gambogenic acid and formed; And

In a preferred embodiment of the present invention, this is used for suppressing the pharmaceutical composition of the growth of a lesion/cancer cell to include one and is selected from following separate part: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E.In a better concrete example of the present invention, this is used for suppressing the pharmaceutical composition of the growth of a lesion/cancer cell to include this separate part TSB-14E.

In addition, applicant is also found to by the activity of cancer-associated proteins matter (cancer-related proteins) or binding ability test: according to this Taiwan Patent case TW I282280 embodiment 1 obtained by the acetone extract product of Gamboge and the separate part TSB-14A to separate part TSB-14E of 5 kinds of acetone extract products that obtains according to the present invention have and suppress cancer-associated proteins matter { such as ring oxygenation-2 (cyclooxygenase-2, COX-2), protein thread amino acid/Soviet Union's amine acid kinase (protein serine/threonine kinase) AURKB (Aurora-B kinases) and CDC2/CCNB1 [cdk1/ cell periodic protein B (cyclin B)], protein tyros kinase (protein tyrosinekinase) ABL1 (ABL), EGF-R ELISA (Epidermal Growth FactorReceptor, EGFR), ERBB2 (HER2), insulin receptor (insulin receptor) and KDR (VEGFR-2), and estrogen receptor alpha (estrogen receptor α, ER α) } activity or the effectiveness of binding ability.Due to these cancer-associated proteins matter and kinds of tumors/cancer [such as colorectal carcinoma (colorectal cancer), cancer of the stomach (gastric cancer), nonsmall-cell lung cancer (non-smallcell lung cancer), the cancer of the brain (brain cancer), thyroid carcinoma (thyroid cancer), nasopharyngeal carcinoma (nasopharyngral cancer), chronic lymphocytic leukemia (chronic myelocyticleukemia), T-cell acute lymphoid blast cell leukemia (T-cell acute lymphoblasticleukemia), lung cancer (lung cancer), colorectal carcinoma (colon cancer), breast cancer (breastcancer), leukemia (leukemia), liver cancer (liver cancer), ovarian cancer (ovarial cancer), renal cancer (renal cancer), cancer of pancreas (pancreas cancer) and carcinoma of endometrium (endometrial cancer)] relevant, applicant is inference accordingly: according to this Taiwan Patent case TWI282280 embodiment 1 obtained by the acetone extract product of Gamboge and the separate part TSB-14A to separate part TSB-14E of 5 kinds of acetone extract products that obtains according to the present invention be available for treatment lesion/cancer disease.

Therefore, the invention provides a kind of pharmaceutical composition being used for the treatment of a lesion/cancer disease, its include following any one:

One acetone extract product of Gamboge; And

One is selected from by the separate part being obtained from this acetone extract product in following formed group: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E as above.

In addition, applicant test by vivo (in vivo) and in vitro (in vitro) and is found to: according to this Taiwan Patent case TW I282280 embodiment 1 obtained by the acetone extract product of Gamboge and the separate part TSB-14A to separate part TSB-14E of 5 kinds of acetone extract products that obtains according to the present invention except Tumor suppression/growth of cancer cells, also there is the activity of pain relieving and anti-inflammation.

Therefore, the invention provides a kind of pharmaceutical composition with analgesic activity, its include following any one:

One acetone extract product of Gamboge; And

In a preferred embodiment of the present invention, this pharmaceutical composition with analgesic activity includes one and is selected from following separate part: separate part TSB-14A, separate part TSB-14B, separate part TSB-14C, separate part TSB-14D and separate part TSB-14E.In a better concrete example of the present invention, this pharmaceutical composition with analgesic activity includes this separate part TSB-14E.

The present invention also provides a kind of pharmaceutical composition with anti-inflammation activity, its include following any one:

One acetone extract product of Gamboge; And

In a preferred embodiment of the present invention, this pharmaceutical composition with anti-inflammation activity includes one and is selected from following separate part: separate part TSB-14A, separate part TSB-14C and separate part TSB-14E.In a better concrete example of the present invention, this pharmaceutical composition with anti-inflammation activity includes this separate part TSB-14E.

Can utilize to have the knack of technology that this skill person knows in detail and be manufactured into one according to pharmaceutical composition of the present invention and be suitable for parenteral tunnel (parenterally), (topically) or oral (orally) formulation of offeing medicine partly, this comprises, but be not limited to, injection product (injection) [such as, aseptic aqueous solution (sterile aqueous solution) or dispersion liquid (dispersion)], aseptic powder (sterile powder), lozenge (tablet), tablet (troche), pill (pill), capsule (capsule) and analogue.

Can offer medicine by a parenteral approach be selected from following formed group according to pharmaceutical composition of the present invention: subcutaneous injection (subcutaneous injection), intramuscular injection (intramuscular injection) and intravenous injection (intravenous injection).

In a preferred embodiment of the present invention, this pharmaceutical composition is made into the formulation being suitable for oral administration medicine supplying.

One can be included further according to pharmaceutical composition of the present invention and be widely used pharmaceutically acceptable supporting agent in medicine producing technology.Such as, this pharmaceutically acceptable supporting agent can comprise one or more and be selected from following reagent: solvent (solvent), emulsifying agent (emulsifier), suspension agent (suspending agent), decomposition agent (decomposer), binding agent (binding agent), vehicle (excipient), tranquilizer (stabilizing agent), sequestrant (chelatingagent), thinner (diluent), jelling agent (gelling agent), sanitas (preservative), lubricant (lubricant), absorption delay agent (absorption delayingagent), fluidizer (plasticizer), weighting agent (filling agent), disintegrating agent (disintegrant), interfacial agent (surfactant), thickening material (thickening agent), liposome (liposome) and analogue.

Dosage according to pharmaceutical composition of the present invention can change depending on following factors with dispensing number of times: the seriousness of the disease that be treated, dosing way, and the body weight of the individuality that will be treated, age, physical appearance and reaction.Generally speaking, according to pharmaceutical composition of the present invention every day dosage normally 2.1mg/Kg body weight to 3.0mg/Kg body weight, in single dose or be divided into the form of several dosage, and by genuine, the oral ground of parenteral or can offer medicine partly.

The present invention will be described further with regard to the following examples, but it is to be understood that these embodiments are for illustrating, and should not be interpreted as the restriction in enforcement of the present invention.

< embodiment >

General operation procedure (General procedures):

The fusing point of product is use one micro melting point determinato (micro melting-pointapparatus) (Yanaco, Japan) and is detected.

Electron bombardment mass spectrum (electron impact mass spectrometry, EIMS) and high electron bombardment mass spectrum (high-resolution electron impact mass spectrometry, HREIMS) of resolving be that use one MAT-95XL height is resolved mass spectrograph and detected.

Fast atom bombardment mass spectrum (fast atom bombardment mass spectrometry, FABMS) and high fast atom bombardment mass spectrum (high-resolution fast atombombardment mass spectrometry, HRFABMS) of resolving be use one JEOL JMSSX SX 102A or Finnigan MAT 95XL mass spectrograph and detected.

1H-NMR and 13C-NMR spectrum is use one BRUKER AM400, VARIAN GEMINI-400 or BRUKER ADVANCE DMX-600 spectrograph (spectrometers) and is detected, and the solvent used is CDCl3 or acetone-d6, and chemical shift (δ) is in units of ppm.

The instrument that AG (analytical) reverse phase high performance energy liquid chromatography (LC) (reversed phase highperformance liquid chromatography, RP-HPLC) uses is as follows: Hitachi L-7400 UV Detector, Hitachi L-7100Pump, Hitachi HPLC D-7000System; Column oven (column oven) is Super CO-150 (Enshine, Taiwan); Protection tubing string (guard column) is SecurityGuard Cartridge C8 (length: 3.0mmx4.0mm) (Phenomenex, USA); Analyzing tubing string is Luna 3 μ C8 (2) (length: 4.6mmx150mm) (Phenomenex, USA).And the operational condition that AG RP-HPLC uses is as follows: Sample injection volume is 5 μ L; Movement is 0.05% trifluoroacetic acid (trifluoroacetic acid, the TFA) aqueous solution/100% acetonitrile (acetonitrile)=35: 65 mutually; Tubing string flow velocity is 0.75mL/min; Column oven temperature is 35 DEG C; Detecting wavelength is set to 360nm.

The instrument that half preparation scale (semipreparative) RP-HPLC uses is as follows: HitachiL-7400UV Detector, Hitachi L-7150Pump, Hitachi HPLC D-7000 System; Column oven is Super CO-150 (Enshine, Taiwan); Protection tubing string is SecurityGuardCartridge C12 (length: 10mmx10mm) (Phenomenex, USA); Analyzing tubing string is Synergi 4 μ C12 (length: 10mmx150mm) (Phenomenex, USA).And the operational condition that half preparation scale RP-HPLC uses is as follows: Sample injection volume is 250 μ L; Mobile mutually depending on experiment needs; Gradient elution (gradient elution) mode needs depending on experiment; Tubing string flow velocity is 4.5mL/min; Column oven temperature is 27 DEG C; Detecting wavelength is set to 360nm.

AG (analytical) the reverse phase high performance energy liquid chromatography (LC) (reversed phase high performanceliquid chromatography, RP-HPLC) of the acetone extract product TSB-14 of embodiment 1. Gamboge is analyzed

This experiment mainly by according to the Taiwan Patent case TW I282280 of applicant (corresponding to US7,138,428 B2 and CN 100413868 C) the acetone extract product TSB-14 of the Gamboge obtained by embodiment 1 bring and carry out AG RP-HPLC, in order to do to include those compositions in the acetone extract product TSB-14 confirming Gamboge further.

The acetone extract product TSB-14 of the Gamboge of 1.0mg is dissolved in the acetone of 1mL, and according to above " general operation procedure " when described in method carry out AG RP-HPLC, and obtain an AG RP-HPLC elution figure (analytical RP-HPLC elutionprofile) as shown in Figure 1.

As seen from Figure 1, the acetone extract product TSB-14 of Gamboge has 35 main elution crests (being denoted as crest 1 to crest 35 respectively) within the residence time (retention time) of the 0th to the 80th minute.Experimental result according to Fig. 1, applicant thinks can isolate 35 kinds of compounds from the acetone extract product TSB-14 of Gamboge, therefore, uses the experiment that the acetone extract product TSB-14 of Gamboge carries out below.

Embodiment 2. preparation is derived from the purified compound of the acetone extract product of Gamboge

The half preparation scale RP-HPLC of the acetone extract product TSB-14 of A, Gamboge:

The acetone extract product TSB-14 of the Gamboge of 3g is dissolved in the acetone/acetonitrile (v/v=1: 9) of 30mL, and according to above " general operation procedure " when described in method carry out half preparation scale RP-HPLC, wherein movement is the 0.05%TFA aqueous solution/65% acetonitrile=35 mutually: 65.The half preparation scale RP-HPLC elution figure obtained is shown in Fig. 2.Can know after Fig. 2 and Fig. 1 of embodiment 1 above are done a comparison and find out the position of crest 1 to crest 35 at Fig. 2.These 35 crests are distinguished into 3 sections (sections) (namely, section 1 to section 3) by applicant, and wherein section 1 is containing crest 1 to crest 12, and the residence time is the 0th to the 42nd minute; Section 2 is containing crest 13 to crest 24, and the residence time is the 42nd to the 135th minute; And section 3 is containing crest 25 to crest 35, the residence time is the 135th to the 280th minute.

The preparation of B, separate part 1 to separate part 3:

The acetone extract product TSB-14 of the Gamboge of 3g is dissolved in the acetone/acetonitrile (v/v=1: 9) of 30mL, and according to above " general operation procedure " when described in method carry out half preparation scale RP-HPLC, wherein movement is the 0.05%TFA aqueous solution/65% acetonitrile=35 mutually: 65.When elution, collect the washings (eluate) corresponding to each section respectively according to section 1 to the section 3 (see Fig. 2) of the half preparation scale RP-HPLC elution figure obtained in the middle of A item above.

Each washings carries out distribution with H2O with ethyl acetate (ethyl acetate) respectively and is separated (partitioning), then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener (vacuum rotatory evaporator), and obtain separate part 1, separate part 2 and separate part 3.

Half preparation scale RP-HPLC of C, separate part 1 to separate part 3:

The separate part 1 to 3 (respectively getting 100mg) obtained in the middle of B item is above dissolved in the acetone of 1mL respectively, and according to B item above when described in method carry out half preparation scale RP-HPLC, and obtain half preparation scale RP-HPLC elution figure (see Fig. 3 to Fig. 5) of separate part 1 to 3.Can know after the crest location of Fig. 3 to Fig. 5 is done a comparison with Fig. 2 institute tool person respectively and find out, half preparation scale RP-HPLC elution figure of separate part 1 to separate part 3 is respectively containing crest 1 to crest 12, crest 13 to crest 24 and crest 25 to crest 35.

D, preparation are derived from the purified compound of separate part 1:

Obtained separate part 1 in the middle of the B item above of 100mg is dissolved in the acetone of 1mL, and according to B item above when described in method carry out half preparation scale RP-HPLC.When elution, washings 1 to the washings 12 corresponding respectively to crest 1 to crest 12 is collected in half preparation scale RP-HPLC elution figure (see Fig. 3) according to the separate part 1 obtained in the middle of C item above.

In addition, in order to obtain a large amount of washings 1 to washingses 12, repeat half above-mentioned preparation scale RP-HPLC 50 to 100 times.Obtained washings 1 to washings 12 is collected in the amber glass bottle of 4L respectively, then carries out concentrated with vacuum revolution thickener and obtain enriched material 1 to enriched material 12.Then, enriched material 1 to enriched material 12 carries out distribution with H2O with ethyl acetate respectively and is separated, and then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener, and obtain crude product Gh-3261, Gh-3271, Gh-3272, Gh-3311, Gh-3332, Gh-1036, Gh-3291, Gh-631, Gh-1052, Gh-3351, Gh-3353 and Gh-3352.

Obtained 12 kinds of crude products (respectively getting 100mg) are dissolved in the acetone of 1.0mL respectively, and respectively according to above " general operation procedure " when described in method carry out AG RP-HPLC, and obtain the AG RP-HPLC elution figure of each crude product.If the AG RP-HPLC elution figure of the crude product obtained does not demonstrate single crest, AG RP-HPLC elution figure remaining crude product repeated half above-mentioned preparation scale RP-HPLC to carry out further purifying, until can demonstrate single crest.After completing purifying, the weight of each product is shown in table 1 below.

Table 1. is derived from the weight of the purified product of separate part 1

Crest is numbered	Products nr	Weight (mg)
			1	Gh-3261	9
2	Gh-3271	8
			3	Gh-3272	7
4	Gh-3311	7
			5	Gh-3332	6
6	Gh-1036	10

7	Gh-3291	7
			8	Gh-631	12
9	Gh-1052	6
			10	Gh-3351	9
11	Gh-3353	7
			12	Gh-3352	5

E, preparation are derived from the purified compound of separate part 2:

Obtained separate part 2 in the middle of the B item above of 100mg is dissolved in the acetone of 1mL, and according to B item above when described in method carry out half preparation scale RP-HPLC.When elution, the washings 13 to 24 corresponding respectively to crest 13 to crest 24 is collected in half preparation scale RP-HPLC elution figure (see Fig. 4) according to the separate part 2 obtained in the middle of C item above.

In addition, in order to obtain a large amount of washings 13 to washingses 24, repeat half above-mentioned preparation scale RP-HPLC 50 to 100 times.Obtained washings 13 to washings 24 is collected in the amber glass bottle of 4L respectively, then carries out concentrated with vacuum revolution thickener and obtain enriched material 13 to enriched material 24.Enriched material 13 to the enriched material 24 obtained connect and according to D item above when described in method carry out distribution isolation and purification.After completing purifying, the weight of each product is shown in table 2 below.

Table 2. is derived from the weight of the purified product of separate part 2

Crest is numbered	Products nr	Weight (mg)
			13	Gh-47	14
14	Gh-4602	48
			15	Gh-4601	27
16	Gh-1601-A	20
			17	Gh-1050	16
18	Gh-1602	16
			19	Gh-1631	19
20	Gh-2641-1	17
			21	Gh-2501	13
22	Gh-2642	19
			23	Gh-2507	8
24	Gh-2505	56

F, preparation are derived from the purified compound of separate part 3:

Obtained separate part 3 in the middle of the B item above of 100mg is dissolved in the acetone of 1mL, and according to B item above when described in method carry out half preparation scale RP-HPLC.When elution, washings 25 to the washings 35 corresponding respectively to crest 25 to crest 35 is collected in half preparation scale RP-HPLC elution figure (see Fig. 5) according to the separate part 3 obtained in the middle of C item above.

In addition, in order to obtain a large amount of washings 25 to washingses 35, repeat half above-mentioned preparation scale RP-HPLC 50 to 100 times.Obtained washings 25 to washings 35 is collected in the amber glass bottle of 4L respectively, then carries out concentrated with vacuum revolution thickener and obtain enriched material 25 to enriched material 35.Enriched material 25 to the enriched material 35 obtained connect and according to D item above when described in method carry out distribution isolation and purification.After completing purifying, the weight of each product is shown in table 3 below.

Table 3. is derived from the weight of the purified product of separate part 3

Crest is numbered	Products nr	Weight (mg)
			25	Gh-2508	18
26	Gh-2603-1	16
			27	Gh-2603-2	18
28	Gh-1641	30
			29	Gh-1642	25
30	Gh-2605	162
			31	Gh-2606	124
32	Gh-2607-B	10
			33	Gh-2607-1A	12
34	Gh-2301	14
			35	Gh-4301	22

The characterized of the compound of embodiment 3. purifying self-separation part 1 to separate part 3

The product of 35 provenance self-separation part 1 to the separate parts 3 obtained in embodiment 2 be above according to above " general operation procedure " when described in method to carry out physics and chemistry analysis, comprise fusing point, 1H-NMR, 13C-NMR, with core association spectrum (homonuclear correlationspectroscopy, 1H-1H COSY), heteronuclear Multiple-quantum association (heteronuclearmultiple-quantum coherence, HMQC), heteronuclear multiple-bond association (heteronuclearmultiple-bond coherence, HMBC), core Ao Fohaose effect spectrum (nuclearOverhauser effect spectroscopy, NOESY), EIMS, HREIMS, FABMS and HRFABMS.The experimental data obtained is summarized as follows:

1. product Gh-47:

Product Gh-47 is as follows by the character recorded:

Yellow plate crystal, fusing point: 204-209 DEG C.

EIMS m/z (relative intensity): 560 [M]+(100), 545 (47), 532 (22), 517 (36), 405 (44), 389 (11), 363 (24), 349 (17), 307 (12), 287 (22), 285 (16), 245 (15), 215 (12), 189 (5).

1H-NMR(400MHz，CDCl3)：δ7.56(1H，d，J＝6.9Hz)，6.63(1H，d，J＝9.6Hz)，5.51(1H，d，J＝9.6Hz)，5.12(1H，dd，J＝13.6，6.8Hz)，3.52(1H，dd，J＝6.2，4.7Hz)，3.26(2H，d，J＝7.4Hz)，2.66(1H，m)，2.55(2H，m)，2.35(1H，dd，J＝13.4，4.6Hz)，1.75(3H，s)，1.72(3H，s)，1.65(3H，s)，1.44(3H，s)，1.43(3H，s)，1.35(3H，s)，1.30(3H，s)。

13C-NMR(100MHz，CDCl3)：δ200.93，178.91，171.86，161.10，157.64，157.28，136.97，135.39，133.35，131.70，128.63，126.18，122.12，115.44，108.27，103.18，100.49，90.71，83.71，83.64，78.65，49.06，46.88，29.95，28.93，28.33，25.71，25.31，21.62，18.09，11.34。

According to measured spectroscopic data, product Gh-47 is identified is a known compound with the following chemical structure formula, namely isomorellic acid (isomorellic acid):

2. product Gh-631:

Product Gh-631 is as follows by the character recorded:

Yellow needles, fusing point: 115-118 DEG C.

EIMS m/z (relative intensity): 394 [M]+(66), 393 (23), 379 (100), 339 (30), 323 (16), 311 (13), 295 (5), 278 (10), 203 (4), 162 (7).

1H-NMR (600MHz, acetone-d6): δ 13.09 (1H, s, OH-1), 7.40 (1H, s, H-8), 6.55 (1H, d, J=10.0Hz, H-11), 6.32 (1H, s, H-2), 5.87 (1H, d, J=10.0Hz, H-12), 5.36 (H, m, H-17), 3.55 (2H, d, J=7.3Hz, H-16), 1.84 (3H, s, H-20), 1.63 (3H, s, H-19), 1.48 (6H, s, H-14, H-15).

13C-NMR (150MHz, acetone-d6): δ 181.24 (C=O), 163.41 (C-3), 162.26 (C-1), 155.56 (C-4a), 146.94 (C-6), 146.43 (C-10a), 134.42 (C-18), 132.30 (C-12), 131.77 (C-5), 123.33 (C-17), 122.08 (C-11), 119.03 (C-7), 115.20 (C-8a), 113.15 (C-8), 107.54 (C-4), 103.26 (C-9a), 98.40 (C-2), 78.84 (C-13), 28.26 (C-14, 15), 25.93 (C-19), 22.16 (C-16), 18.00 (C-20).

According to measured spectroscopic data, product Gh-631 is identified is a known compound with the following chemical structure formula, namely formoxanthone A (formoxanthone A):

3. product Gh-4601:

Product Gh-4601 is as follows by the character recorded:

Orange powder, fusing point: 137-139 DEG C.

EIMS m/z (relative intensity): 546 [M]+(100), 531 (18), 518 (44), 503 (40), 485 (9), 433 (7), 405 (33), 391 (10), 363 (19), 349 (13), 307 (10), 287 (25), 245 (8), 231 (18), 214 (12), 189 (5), 105 (6).

1H-NMR (400MHz, CDCl3): δ 12.68 (1H, s), 7.41 (1H, d, J=7.2Hz), 6.59 (1H, d, J=10.0Hz), 5.49 (1H, d, J=10.0Hz), 5.19 (1H, t, J=7.0Hz), 4.73 (1H, t, J=8.0Hz), 3.61 (2H, q, J=10.2Hz), 3.49 (1H, d, J=4.7Hz), 3.47 (1H, d, J=4.7Hz), 3.32 (1H, dd, J=14.5, 6.8Hz), 3.24 (1H, dd, J=14.3, 7.7Hz), 2.60 (1H, d, J=7.7Hz), 2.48 (1H, d, J=9.4Hz), 2.31 (1H, dd, J=13.5, 4.7Hz), 1.74, 1.68, 1.64, 1.25, 1.01 (each 3H, s), 1.41 (6H, s).

13C-NMR(100MHz，CDCl3)：δ203.35，180.33，161.06，157.80，157.44，138.01，134.40，133.63，131.87，126.31，121.90，118.20，115.50，108.44，103.00，100.72，90.48，84.50，83.41，78.66，67.92，49.09，47.00，30.09，28.96，28.28，28.21，25.71，25.29，21.57，18.14，12.49。

According to measured spectroscopic data, product Gh-4601 is identified is a known compound with the following chemical structure formula, namely isomorellinol (isomorellinol):

4. product Gh-4602:

Product Gh-4602 is as follows by the character recorded:

Orange powder, fusing point: 106-109 DEG C.

EIMS m/z (relative intensity): 560 [M]+(100), 545 (56), 532 (63), 517 (48), 487 (12), 433 (9), 405 (81), 391 (22), 363 (38), 349 (24), 307 (18), 287 (64), 245 (40), 231 (21), 215 (20), 189 (10).

1H-NMR (400MHz, CDCl3): δ 12.71 (1H, s), 7.51 (1H, d, J=6.8Hz), 6.49 (1H, d, J=10.0Hz), 6.05 (1H, t, J=7.0Hz), 5.39 (1H, d, J=10.0Hz), 4.99 (1H, d, J=6.0Hz), 3.45 (1H, dd, J=6.4, 4.7Hz), 3.27 (1H, m), 3.08 (1H, m), 2.97 (2H, s ept, J=8.0Hz), 2.49 (1H, d, J=9.3Hz), 2.28 (1H, dd, J=13.4, 4.5Hz), 1.70, 1.69, 1.67, 1.60, 1.36, 1.34, 1.26 (each 3H, s).

13C-NMR(100MHz，CDCl3)：δ203.47，179.07，171.74，161.22，157.65，157.34，138.46，135.39，133.42，131.46，127.64，126.00，122.22，115.44，108.04，103.16，100.55，90.93，83.82，78.55，49.01，46.80，29.87，29.26，28.82，28.40，28.20，25.68，25.14，21.57，20.63，18.06。

According to measured spectroscopic data, product Gh-4602 is identified is a known compound with the following chemical structure formula, namely morellic acid (morellic acid):

5. product Gh-2301:

Product Gh-2301 is as follows by the character recorded:

Orange needle-like crystals, fusing point: 109-110 DEG C.

EIMS m/z (relative intensity): 530 [M]+(100), 515 (22), 502 (92), 488 (30), 487 (83), 459 (11), 433 (20), 405 (49), 391 (15), 363 (24), 349 (16), 307 (13), 287 (27), 231 (13), 215 (37), 189 (6).

1H-NMR (400MHz, CDCl3): δ 12.89 (1H, s), 7.45 (1H, d, J=7.2Hz), 7.27 (1H, s), 6.65 (1H, d, J=10.4Hz), 5.53 (1H, d, J=9.6Hz), 5.30 (1H, br d, J=6.0Hz), 4.43 (1H, br s), 3.49 (1H, m), 3.32 (2H, m), 2.50 (2H, m), 2.34 (1H, m), 1.78,1.71,1.68,1.59,1.33,1.03 (each 3H, s), 1.45 (6H, s).

According to measured spectroscopic data, product Gh-2301 is identified is a known compound with the following chemical structure formula, namely desoxymorellin (desoxymorellin):

6. product Gh-4301:

Product Gh-4301 is as follows by the character recorded:

Yellow gummy solid, fusing point: 85-88 DEG C.

EIMS m/z (relative intensity): 600 [M]+(100), 572 (77), 557 (21), 531 (10), 503 (60), 475 (26), 449 (33), 393 (9), 357 (12), 351 (26), 323 (9), 309 (16), 295 (45), 281 (12), 253 (53), 231 (18), 215 (40), 189 (10), 177 (18), 173 (12), 69 (65).

1H-NMR(600MHz，CDCl3)：δ12.92(1H，s)，7.41(1H，d，J＝7.0Hz)，6.45(1H，s)，5.19(2H，m)，5.03(1H，m)，4.39(1H，m)，3.45(1H，dd，J＝6.9，4.5Hz)，3.36(4H，m)，2.52(2H，m)，2.43(1H，d，J＝9.4Hz)，2.30(1H，dd，J＝13.5，4.7Hz)，2.05(4H，m)，1.78(3H，s)，1.74(3H，s)，1.69(3H，s)，1.654(3H，s)，1.645(3H，s)，1.56(3H，s)，1.33(3H，s)，1.30(1H，m)，1.25(3H，s)，0.98(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.55，179.65，162.98，160.12，156.29，139.04，134.91，133.88，133.80，133.75，131.88，123.74，121.86，121.33，117.76，107.13，106.39，100.70，90.12，84.50，83.11，49.06，46.90，39.66，30.03，29.02，28.78，26.32，25.70，25.63，25.49，25.40，22.05，21.11，18.01，17.63，16.68，16.19。

According to measured spectroscopic data, product Gh-4301 is identified is a known compound with the following chemical structure formula, namely desoxygambogenin (desoxygambogenin):

7. product Gh-2605:

Product Gh-2605 is as follows by the character recorded:

Yellow powder, fusing point: 205-208 DEG C.

EIMS m/z (relative intensity): 628 [M]+(22), 600 (15), 545 (100), 517 (23), 473 (17), 431 (7), 389 (9), 355 (10), 295 (5), 271 (7), 245 (14), 215 (23), 189 (11), 69 (9).

1H-NMR(600MHz，CDCl3)：δ12.73(1H，s)，7.52(1H，d，J＝6.9Hz)，6.57(1H，d，J＝10.2Hz)，6.08(1H，d t，J＝7.5，1.3Hz)，5.35(1H，d，J＝10.2Hz)，5.01(2H，m)，3.46(1H，dd，J＝6.8，4.6Hz)，3.27(1H，dd，J＝14.7，8.1Hz)，3.12(1H，br dd，J＝14.7，5.3Hz)，2.93(2H，t，J＝7.3Hz)，2.49(1H，d，J＝9.3Hz)，2.29(1H，dd，J＝13.4，4.7Hz)，1.98(2H，m)，1.72(3H，d，J＝1.3Hz)，1.71(1H，m)，1.70(3H，s)，1.67(3H，s)，1.62(3H，s)，1.60(3H，s)，1.56(1H，m)，1.52(3H，s)，1.35(3H，s)，1.34(1H，m)，1.27(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.28，178.85，170.88，161.49，157.55，157.35，137.90，135.31，133.34，131.77，131.49，127.70，124.49，123.83，122.25，115.87，107.60，102.73，100.44，90.91，83.92，83.78，81.28，49.00，46.81，41.97，29.85，29.26，28.84，27.69，25.64，25.62，25.16，22.73，21.60，20.73，18.06，17.60。

According to measured spectroscopic data, product Gh-2605 is identified is a known compound with the following chemical structure formula, namely gambogic acid (gambogic acid):

8. product Gh-2606:

Product Gh-2606 is as follows by the character recorded:

Yellow powder, fusing point: 88-92 DEG C.

EIMS m/z (relative intensity): 628 [M]+(24), 600 (11), 545 (100), 517 (21), 473 (12), 431 (4), 389 (8), 347 (6), 245 (7), 215 (14), 189 (5), 69 (4).

1H-NMR(600MHz，CDCl3)：δ12.74(1H，s)，7.53(1H，d，J＝6.9Hz)，6.56(1H，d，J＝10.0Hz)，6.10(1H，d t，J＝7.4，1.2Hz)，5.38(1H，d，J＝10.0Hz)，5.07(1H，br t，J＝7.1Hz)，5.01(1H，br t，J＝6.3Hz)，3.45(1H，dd，J＝6.5，4.9Hz)，3.29(1H，dd，J＝14.6，8.3Hz)，3.13(1H，br dd，J＝14.6，4.3Hz)，2.94(1H，dd，J＝16.1，7.7Hz)，2.87(1H，dd，J＝16.3，6.3Hz)，2.49(1H，d，J＝9.3Hz)，2.28(1H，dd，J＝13.4，4.7Hz)，2.05(2H，m)，1.75(1H，m)，1.72(3H，s)，1.71(3H，s)，1.68 (3H，s)，1.64(3H，s)，1.62(1H，m)，1.61(3H，s)，1.55(3H，s)，1.35(1H，m)，1.31(3H，s)，1.26(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.25，178.92，170.68，161.33，157.59，157.33，137.73，135.51，133.21，131.99，131.39，127.76，124.76，123.82，122.24，115.88，107.79，102.88，100.51，90.98，83.86，83.67，81.10，49.00，46.82，41.69，29.91，29.26，28.78，26.91，25.67，25.60，25.20，22.73，21.61，20.71，18.12，17.58。

According to measured spectroscopic data, product Gh-2606 is identified is a known compound with the following chemical structure formula, namely epigambogic acid (epigambogic acid):

9. product Gh-1641:

Product Gh-1641 is as follows by the character recorded:

Yellow powder, fusing point: 53-56 DEG C.

EIMS m/z (relative intensity): 628 [M]+(32), 600 (6), 545 (100), 517 (12), 473 (6), 431 (2), 389 (4), 355 (5), 245 (8), 215 (11), 189 (3), 69 (5).

1H-NMR(600MHz，CDCl3)：δ12.75(1H，s)，7.53(1H，d，J＝6.9Hz)，6.65(1H，d，J＝10.1Hz)，6.61(1H，t，J＝7.5Hz)，5.41(1H，d，J＝10.1Hz)，5.09(1H，t，J＝6.9Hz)，5.04(1H，t，J＝7.8Hz)，3.49(1H，dd，J＝6.7，4.6Hz)，3.24(2H，m)，2.63(1H，dd，J＝15.6，8.2Hz)，2.53(1H，m)，2.51(1H，d，J＝9.3Hz)，2.32(1H，dd，J＝13.5，4.7Hz)，2.02(2H，dd，J＝15.8，7.7Hz)，1.76(1H，m)，1.70(3H，s)，1.69(3H，s)，1.63(3H，s)，1.62(3H，s)，1.59(1H，m)，1.53(3H，s)，1.38(3H，s)，1.37(1H，m)，1.34(3H，s)，1.28(3H，s)。

13C-NMR(150MHz，CDCl3)：δ202.97，178.81，171.55，161.41，157.61，157.35，136.98，135.31，133.36，131.81，131.75，128.56，124.77，123.82，122.18，115.93，107.92，102.82，100.40，90.71，83.73，83.65，81.33，49.05，46.87，41.92，29.93，29.05，28.95，27.49，25.67，25.61，25.33，22.75，21.61，18.08，17.59，11.39。

According to measured spectroscopic data, product Gh-1641 is identified is a known compound with the following chemical structure formula, namely Isogambogic acid (isogambogic acid):

10. product Gh-1642:

Product Gh-1642 is as follows by the character recorded:

Yellow powder, fusing point: 55-60 DEG C.

EIMS m/z (relative intensity): 628 [M]+(63), 600 (13), 545 (100), 517 (17), 473 (17), 431 (8), 389 (7), 355 (7), 245 (8), 215 (10), 189 (3), 69 (18).

1H-NMR(600MHz，CDCl3)：δ12.74(1H，s)，7.52(1H，d，J＝7.1Hz)，6.66(1H，d，J＝10.2Hz)，6.50(1H，t，J＝7.9Hz)，5.44(1H，d，J＝10.2Hz)，5.12(1H，t，J＝6.9Hz)，5.06(1H，t，J＝7.8Hz)，3.49(1H，dd，J＝6.8，4.5Hz)，3.25(2H，m)，2.61(2H，m)，2.49(1H，d，J＝9.5Hz)，2.32(1H，dd，J＝13.3，4.7Hz)，2.06(2H，m)，1.78(1H，m)，1.73(3H，s)，1.70(3H，s)，1.64(3H，s)，1.63(3H，s)，1.62(1H，m)，1.55(3H，s)，1.36(3H，s)，1.34(1H，m)，1.28(3H，s)，1.23(3H，s)。

13C-NMR(150MHz，CDCl3)：δ202.92，178.88，170.95，161.32，157.63，157.36，136.75，135.31，133.33，131.97，131.74，128.81，124.81，123.76，122.14，115.96，107.92，102.93，100.49，90.64，83.67，83.62，81.29，49.07，46.95，41.88，30.03，29.06，28.98，27.32，25.71，25.64，25.48，22.75，21.65，18.16，17.62，11.43。

According to measured spectroscopic data, product Gh-1642 is identified is a known compound with the following chemical structure formula, namely epiisogambogic acid (epiisogambogic acid):

11. product Gh-2603-2:

Product Gh-2603-2 is as follows by the character recorded:

Yellow powder, fusing point: 131-135 DEG C.

EIMS m/z (relative intensity): 628 [M]+(92), 600 (91), 545 (54), 517 (46), 474 (100), 473 (88), 459 (18), 431 (29), 417 (15), 391 (33), 355 (37), 349 (25), 295 (18), 253 (21), 245 (25), 215 (30), 189 (18), 69 (20).

1H-NMR(600MHz，CDCl3)：δ12.62(1H，s，OH-6)，7.47(1H，d，J＝6.9Hz，H-10)，6.00(1H，dd，J＝7.6，1.3Hz，H-27)，5.04(1H，t，J＝6.9Hz，H-32)，4.53(1H，s，H1-40)，4.19(1H，s，H2-40)，3.43(1H，dd，J＝13.4，4.6Hz，H-11)，3.40(1H，br s，H-4)，3.23(1H，dd，J＝14.5，8.1Hz，H1-31)，3.10(1H，dd，J＝14.5，5.6Hz，H2-31)，2.94(2H，d，J＝7.5Hz，H-26)，2.51(1H，d，J＝9.3Hz，H-22)，2.28(1H，dd，J＝13.4，4.7Hz，H1-21)，2.08(1H，br d，J＝12.6Hz，H-37)，1.88(1H，br d，J＝13.6Hz，H1-20)，1.82(3H，s，H-39)，1.77(1H，dd，J＝13.1，2.8Hz，H1-3)，1.71(1H，m，H2-3)，1.70(3H，s，H-34)，1.69(3H，s，H-25)，1.68(3H，s，H-29)，1.62(3H，s，H-35)，1.52(1H，dt，J＝13.4，4.9Hz，H2-20)，1.36(1H，m，H1-36)，1.33(1H，m，H2-21)，1.30(3H，s，H-19)，1.29(1H，m，H2-36)，1.27(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ203.68(C-12)，178.47(C-8)，171.33(C-30)，164.44(C-18)，160.77(C-6)，155.27(C-16)，148.87(C-38)，138.06(C-27)，134.30(C-10)，133.93(C-9)，131.07(C-33)，127.77(C-28)，122.44(C-32)，108.60(C-40)，106.26(C-17)，104.14(C-5)，99.57(C-7)，90.42(C-14)，84.20(C-23)，83.93(C-13)，77.05(C-2)，48.91(C-22)，48.10(C-37)，46.73(C-11)，39.27(C-20)，36.52(C-3)，29.90(C-25)，29.18(C-26)，28.85(C-4)，28.85(C-24)，28.37(C-19)，25.74(C-35)，25.18(C-21)，22.96(C-39)，22.78(C-36)，21.84(C-31)，20.71(C-29)，18.14(C-34)。

The EIMS data presentation of product Gh-2603-2: have a part peak [M]+position at m/z 628, tool person is identical for this and gambogic acid (gambogic acid) institute.In addition, also tool person is identical with known gambogellic acid (gambogellic acid) institute for 1H-NMR and the 13C-NMR spectrum of product Gh-2603-2, and via same core association spectrum (homonuclear correlation spectroscopy, 1H-1H COSY), heteronuclear Multiple-quantum association (heteronuclear multiple-quantumcoherence, HMQC) (J=150Hz) and heteronuclear multiple-bond associate (heteronuclearmultiple-bond coherence, HMBC) (J=8Hz) and are identified errorless.

Also find from the NOESY spectroscopic data of product Gh-2603-2: δ 7.47 (H-10) is associated with δ 3.43 (H-11); δ 3.43 (H-11) is associated with δ 2.28 (H1-21); δ 1.33 (H2-21) is associated with δ 2.51 (H-22); δ 2.28 (H1-21) and δ 2.51 (H-22) is all associated with δ 1.27 (H-24); δ 6.00 (H-27) is associated with δ 1.68 (H-29).This proves that the three-dimensional arrangement (stereostructure) of this part is identical with gambogic acid institute tool person, be all 11S, 13R, 14S, 22S configuration (configuration), and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.

In the monoterpene part (monoterpene moiety) of gambogellic acid, be affixed to methine protons (the methine proton attached to the isopropenylgroup) [δ 2.08 (1H of lsopropenyl group, br d, J=12.6Hz, H-37) Signal aspects]: the methine protons and the neighbour proton (vicinal proton) that are affixed to lsopropenyl group have sizable neighbour coupling constant (vicinal coupling constant) (J=12.6Hz), they are couplings of axial-axial (axial-axial), so H-37 is axial configuration.

In addition, also find from NOESY spectroscopic data: δ 3.40 (H-4), except being associated with δ 2.08 (H-37), is also associated with δ 1.77 (H1-3) and δ 1.71 (H2-3); δ 1.77 (H1-3) is associated with δ 1.30 (H-19); δ 2.08 (H-37) is associated with δ 1.77 (H1-3) and δ 1.52 (H2-20).δ 3.40 (H-4) is associated with δ 1.68 (H-29), so C-2 is R configuration.The monoterpene part (monoterpene moiety) of product Gh-2603-2 is the chair form configuration (rigid chair conformation) of rigidity, and H-37 and H-3 and H-20 has 1,3-bis-axle interaction (1,3-diaxial interaction), and the methyl group (H-19) be connected on C-2 position is all equator to (equatorial), so the three-dimensional arrangement of this part is 2R, 4R, 37S with the lsopropenyl group (isopropenyl group) be connected on C-37 position.

Comprehensive above data, product Gh-2603-2 is identified is a known compound with the following chemical structure formula, namely gambogellic acid (gambogellic acid):

12. product Gh-2603-1:

Product Gh-2603-1 is as follows by the character recorded:

Yellow powder, fusing point: 115-120 DEG C.

EIMS m/z (relative intensity): 628 [M]+(92), 600 (100), 545 (68), 517 (49), 474 (98), 473 (80), 459 (17), 431 (25), 417 (13), 391 (28), 355 (24), 349 (18), 295 (11), 253 (12), 245 (13), 215 (16), 189 (7), 69 (15).

1H-NMR(600MHz，CDCl3)：δ12.68(1H，s，OH-6)，7.48(1H，d，J＝6.9Hz，H-10)，5.99(1H，dt，J＝8.0，1.3Hz，H-27)，5.00(1H，br t，J＝6.7Hz，H-32)，4.52(1H，s，H1-40)，4.26(1H，s，H2-40)，3.50(1H，brs，H-4)，3.44(1H，dd，J＝6.8，4.5Hz，H-11)，3.31(1H，dd，J＝14.6， 8.2Hz，H1-31)，3.20(1H，br dd，J＝14.5，3.8Hz，H2-31)，3.01(1H，dd，J＝16.5，7.6Hz，H1-26)，2.84(1H，ddd，J＝16.6，7.1，1.4Hz，H2-26)，2.46(1H，d，J＝9.4Hz，H-22)，2.29(1H，dd，J＝13.5，4.6Hz，H1-21)，2.13(1H，br d，J＝12.4Hz，H-37)，1.92(1H，br dd，J＝13.8，2.0Hz，H1-20)，1.86(1H，dd，J＝6.6，2.7Hz，H1-3)，1.84(3H，s，H-39)，1.70(1H，m，H2-3)，1.695(3H，s，H-34)，1.69(6H，s，H-25，H-29)，1.61(3H，s，H-35)，1.54(1H，dt，J＝13.5，5.2Hz，H2-20)，1.36(2H，m，H-36)，1.34(1H，m，H2-21)，1.33(3H，s，H-19)，1.27(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ203.23(C-12)，178.53(C-8)，170.69(C-30)，164.50(C-18)，160.92(C-6)，155.39(C-16)，147.30(C-38)，137.10(C-27)，134.59(C-10)，133.70(C-9)，131.11(C-33)，128.00(C-28)，122.59(C-32)，109.28(C-40)，106.34(C-17)，104.24(C-5)，99.48(C-7)，90.48(C-14)，83.90(C-23)，83.61(C-13)，77.09(C-2)，49.13(C-22)，48.32(C-37)，46.74(C-11)，39.33(C-20)，37.13(C-3)，29.89(C-25)，29.50(C-26)，29.09(C-4)，28.95(C-24)，28.52(C-19)，25.69(C-35)，25.41(C-21)，22.93(C-39)，22.69(C-36)，21.90(C-31)，20.56(C-29)，18.16(C-34)。

The EIMS data presentation of product Gh-2603-1: have a part peak [M]+position at m/z 628, cataclasm graphic (fragmentationpatterns) of this and product Gh-2603-2 (namely, gambogellic acid) is identical.In addition, 1H-NMR and the 13C-NMR spectrum of product Gh-2603-1 is similar to product Gh-2603-2 institute tool person haply.

The 1H-NMR spectrum display of product Gh-2603-1: the proton signal being connected to the side chain on C-13 position is different from product Gh-2603-2 institute tool person, methene proton (methylene proton) signal in 2-methyl-2-butenoic acid (2-methyl-2-butenoic acid) is by originally simple doublet (doublet) [δ 2.94 (2H, d, J=7.5Hz, H-26) δ 3.01 (1H] is become, dd, J=16.5,7.6Hz, H1-26) and δ 2.84 (1H, ddd, J=16.6,7.1,1.4Hz, H2-26) signal.This is because be that in the structure of the gambogellic acid of R configuration, methylene group (C-26) can rotate freely (free rotation) at C-2.If C-2 is S configuration, then formed monoterpene p-alkene in Meng ring (monoterpene p-menthene ring) and lsopropenyl group (isopropenyl group) can form steric barrier (steric hindrance) and impact rotates freely, and then make 2 proton non-equivalences (nonequivalence) of methylene group, and anisotropy effect (anisotropic effect) is produced to relevant proton.In addition, the methine protons δ 2.13 (1H, br d, J=12.4Hz, H-37) being affixed to lsopropenyl group has large coupling constant (J=12.4Hz), and this represents that methine protons is axially.

The structure of product Gh-2603-1 is identified via 1H-1H COSY, HMQC and HMBC spectrum, wherein about the HMBC spectroscopic data display of p-ene-type in Meng monoterpene outer-methene proton (exo-methylene proton) [δ 4.52 (H1-40) and δ 4.26 (H2-40)] is associated with δ 22.93 (C-39) and δ 48.32 (C-37); C-3 methene proton [δ 1.86 (H1-3)] is associated with δ 104.24 (C-5), δ 29.09 (C-4), δ 48.32 (C-37) and δ 147.30 (C-38); C-20 methene proton [δ 1.92 (H1-20)] is associated with δ 48.32 (C-37) and δ 29.09 (C-4).In addition, the mesomethylene carbon [δ 37.13 (C-3) and δ 39.33 (C-20)] being connected to methyl proton [δ 1.33 (H-19)] on C-2 position adjacent with 2 is associated.

Also find from the HMBC spectroscopic data of product Gh-2603-1: H-3 is associated with the C-5 of aromatic nucleus, and the lsopropenyl group be connected on C-37 position and the methyl proton (H-19) be connected on C-2 position be all equator to.Therefore, the structure of monoterpene part is the chair form configuration of rigidity.

In addition, find from the NOESY spectroscopic data of product Gh-2603-1: δ 2.13 (H-37) is associated with δ 1.36 (H-36) and δ 1.54 (H2-20).This represents that axial hydrogen H-37 has 1 with the axial hydrogen (H-3 and H-20) adjacent to 2 methylene groups of C-2,3-bis-axle interaction, so the three-dimensional arrangement of p-alkene in the Meng (p-menthene) is 2S, 4S, 37R, this is contrary with product Gh-2603-2 institute tool person.In addition, also find from NOESY spectroscopic data: δ 7.48 (H-10) is associated with δ 3.44 (H-11); δ 3.44 (H-11) is associated with δ 2.29 (H1-21); δ 1.34 (H2-21) is associated with δ 2.46 (H-22); δ 2.29 (H1-21) and δ 2.46 (H-22) is all associated with δ 1.27 (H-24); δ 5.99 (H-27) is associated with δ 1.69 (H-29).This proves that the three-dimensional arrangement of r-piperazine click (r-pyrone) right half part of product Gh-2603-1 is identical with gambogic acid or gambogellic acid institute tool person, be all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are for trans, and double bond Δ 27,28 is Z configuration.

Comprehensive above data, product Gh-2603-1 is identified is a compounds with the following chemical structure formula:

Product Gh-2603-1 is named as epigambogellic acid (epigambogellic acid), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10R, 13S, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxyl-3, 3, 13-trimethylammonium-15-(3-methyl-2-butene base)-10-(1-methyl ethylene)-7, 18-dioxy-1, 5:9, 13-dimethylene-1H, 3H, 9H-furo [3.4-g] oxygen is [3.2-b]-1-base also]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10R, 13S, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxy-3, 3, 13-trimethyl-15-(3-methyl-2-butenyl)-10-(1-methylethenyl)-7, 18-dioxo-1, 5:9, 13-dimethano-1H, 3H, 9H-furo [3.4-g] oxocino [3.2-b] xanthen-1-yl]-, (2Z)-].

13. product Gh-2607-B:

Product Gh-2607-B is as follows by the character recorded:

Yellow powder, fusing point: 120-125 DEG C.

EIMS m/z (relative intensity): 628 [M]+(33), 600 (17), 545 (100), 517 (23), 499 (4), 474 (14), 431 (5), 389 (8), 355 (7), 347 (6), 245 (5), 215 (9), 189 (3), 69 (4).HREIMS [M]+m/z:628.3034; Calculated value about C38H44O8: 628.3036.

1H-NMR(600MHz，CDCl3)：δ12.57(1H，s，OH-6)，7.50(1H，d，J＝6.9Hz，H-10)，5.86(1H，dt，J＝7.6，1.2Hz，H-27)，5.10(1H，br t，J＝7.0Hz，H-32)，3.46(1H，dd，J＝6.8，4.5Hz，H-11)，3.28(1H，dd，J＝14.7，8.5Hz，H1-31)，3.15(1H，m，H1-26)，3.12(1H，m，H2-31)，2.95(1H，d，J＝9.6Hz，H-3)，2.91(1H，ddd，J＝15.9，6.9，1.4Hz，H2-26)，2.50(1H，d，J＝9.3Hz，H-22)，2.42(1H，dd，J＝9.5，7.4Hz，H-4)，2.31(1H，m，H-37)，2.29(1H，m，H1-21)，1.74(1H，m，H1-20)，1.71(3H，s，H-34)，1.69(3H，s，H-25)，1.67(3H，s，H-29)，1.60(3H，s，H-35)，1.60(1H，m，H1-36)，1.53(1H，m，H2-20)，1.49(1H，m，H2-36)，1.36(1H，m，H2-21)，1.30(3H，s，H-40)，1.28(3H，s，H-19)，1.27(3H，s，H-24)，0.71(3H，s，H-39)。

13C-NMR(150MHz，CDCl3)：δ203.71(C-12)，178.89(C-8)，171.22(C-30)，161.53(C-18)，161.17(C-6)，155.04(C-16)，137.79(C-27)，134.63(C-10)，133.88(C-9)，130.85(C-33)，128.20(C-28)，122.37(C-32)，108.88(C-17)，105.32(C-5)，100.38(C-7)，90.33(C-14)，85.18(C-2)，84.18(C-23)，83.86(C-13)，48.94(C-22)，46.81(C-11)，46.23(C-37)，38.81(C-38)，38.58(C-20)，36.93(C-4)，35.01(C-3)，33.45(C-40)，29.91(C-25)，29.25(C-26)，28.93(C-24)，27.35(C-19)，25.74(C-35)，25.61(C-36)，25.32(C-21)，21.83(C-31)，20.78(C-29)，18.19(C-34)，17.68(C-39)。

The EIMS data presentation of product Gh-2607-B: have a part peak [M]+position at m/z 628, and HREIMS data presentation [M]+m/z 628.3034, this represents that the molecular formula of product Gh-2607-B is identical with above-mentioned gambogic acid, gambogellic acid and their epimer (epimers) institute tool person, is all C38H44O8.

1H-NMR and the 13C-NMR spectrum display of product Gh-2607-B: do not have two replacement double bonds (disubstituted double bond) in the piperazine of product Gh-2607-B mutters ring (pyran ring), also do not have the terminal double link (terminal double bond) of lsopropenyl group.

In addition, undertaken confirming rear discovery by 1H-1H COSY, HMQC and HMBC spectrum, product Gh-2607-B is except monoterpene part (monot erpene moiety), and other parts are all similar to gambogellic acid institute tool person.With gambogellic acid in comparison, product Gh-2607-B has lacked a double bond, thus infers a product Gh-2607-B structurally ring more than gambogellic acid.

The 1H-1H COSY spectroscopic data display of product Gh-2607-B: the methine protons [δ 2.95 (1H of 1 doublet, d, J=9.6Hz, H-3)] only with another methine protons [δ 2.42 (1H, dd, J=9.5,7.4Hz, H-4)] coupling, and the latter and 1 methine protons [δ 2.31 (1H, m, H-37)] coupling; δ 2.31 (1H, m, H-37) is coupled with 2 methene protons [δ 1.60 (1H, m, H1-36) and δ 1.49 (1H, m, H2-36)] of C-36; Another methene proton [δ 1.74 (1H, m, H1-20)] is coupled with 2 methene protons [δ 1.60 (1H, m, H1-36) and δ 1.49 (1H, m, H2-36)] of C-36.This proves that the monoterpene part of product Gh-2607-B still has 2 adjacent methylene groups [C-36 (δ 25.61) and C-20 (δ 38.58)] and 3 methin groups [C-3 (δ 35.01), C-4 (δ 36.93) and C-37 (δ 46.23)].The methin groups (C-3) of product Gh-2607-B is the conversion (transformation) of the methylene group (C-3) of gambogellic acid, this conversion is form bond by C-3 and C-38 and instead of a terminal double link (terminal double bond) through inferring, and then form the structure of pinane type monoterpene (pinane typemonoterpene), this structure contains 1 tetramethylene (cyclobutane), 1 together with-dimethyl group (gem-dimethyl group) [C-39 (δ 17.68) and C-40 (δ 33.45)], 1 contains oxygen level Four carbon (oxygen-bearing quaternary carbon) [C-2 (δ 85.18)] and is affixed to this three grades of methyl groups (the tertiary methyl groupattached to the oxygen-bearing quaternary carbon) [C-19 (δ 27.35)] containing oxygen level Four carbon.

Find from the HMBC spectroscopic data of product Gh-2607-B: δ 2.95 (H-3), except being associated with aromatic nucleus carbon δ 105.32 (C-5), δ 161.17 (C-6) and δ 161.53 (C-18), is also associated with δ 85.18 (C-2), δ 46.23 (C-37), δ 38.81 (C-38), δ 38.58 (C-20), δ 36.93 (C-4), δ 33.45 (C-40) and δ 17.68 (C-39); δ 2.42 (H-4) is associated with δ 105.32 (C-5), δ 85.18 (C-2), δ 38.81 (C-38), δ 35.01 (C-3), δ 27.35 (C-19) and δ 25.61 (C-36); δ 2.31 (H-37) is associated with δ 85.18 (C-2), δ 38.81 (C-38), δ 35.01 (C-3), δ 36.93 (C-4) and δ 33.45 (C-40).Pinane type monoterpene (the pinane type monoterpene) structure had by the known product Gh-2607-B of above HMBC spectroscopic data links with the C-18 of ehter bond and aromatic nucleus in C-2 position, and C-4 and C-5 is connected, and the methyl group (C-19) be connected on C-2 position be equator to.

Find from the NOESY spectroscopic data of product Gh-2607-B: δ 2.95 (H-3) is associated with δ 2.42 (H-4), δ 1.28 (H-19) and δ 1.30 (H-40); δ 2.42 (H-4) is associated with δ 2.31 (H-37) and δ 1.30 (H-40); δ 2.31 (H-37) is associated with δ 1.30 (H-40); δ 0.71 (H-39) is associated with δ 1.30 (H-40), δ 1.28 (H-19), δ 1.74 (H1-20) and δ 1.53 (H2-20); δ 1.67 (H-29) is associated with δ 5.86 (H-27), but there is no with δ 2.42 (H-4) and δ 2.31 (H-37) and intersect peak (cross peak).This display C-2 is R configuration, methyl group (C-19) for α-equator to, and in pinane (pinane) structure, 3 methine protons H-3, H-4 and H-37 on tetramethylene ring (cyclobutane ring) position are cis (cis), and its three-dimensional arrangement is 3S, 4S, 37R configuration.

In addition, also find from the NOESY spectroscopic data of product Gh-2607-B: δ 7.50 (H-10) is associated with δ 3.46 (H-11); δ 3.46 (H-11) is associated with δ 2.29 (H1-21); δ 1.36 (H2-21) is associated with δ 2.50 (H-22); δ 2.50 (H-22) is associated with δ 1.69 (H-25).This proves that the three-dimensional arrangement of this part is 11S, 13R, 14S, 22S configuration.In addition, because δ 1.67 (H-29) is associated with δ 5.86 (H-27), so H-27 and carboxylic group (C-30) are for trans, double bond Δ 27,28 is Z configuration.

Comprehensive above data, product Gh-2607-B is identified is a compounds with the following chemical structure formula:

Product Gh-2607-B is named as formoxanthone B (formoxanthone B), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10S, 12R, 15R, 18aS)-3a, 4, 5, 7, 12, 13, 14, 15-octahydro-8-hydroxyl-3, 3, 11, 11, 15-pentamethyl--17-(3-methyl-2-butene base)-7, 19-dioxy-1, 5-methylene radical-1H, 3H, 9H-furo [3.4-g]-16-oxygen-ginseng ring [4.4.0.09, 12] decyl also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10S, 12R, 15R, 18aS)-3a, 4, 5, 7, 12, 13, 14, 15-octahydro-8-hydroxy-3, 3, 11, 11, 15-pentamethyl-17-(3-methyl-2-but enyl)-7, 19-dioxo-1, 5-methano-1H, 3H, 9H-furo [3.4-g]-16-oxa-tricyclo [4.4.0.09, 12] decano [3.2-b] xanthen-1-yl]-, (2Z)-].

14. product Gh-2607-1A:

Product Gh-2607-1A is as follows by the character recorded:

Yellow powder, fusing point: 146-151 DEG C.

EIMS m/z (relative intensity): 628 [M]+(27), 601 (13), 545 (100), 517 (20), 473 (12), 389 (8), 355 (7), 347 (7), 245 (10), 215 (18), 189 (8), 69 (11).HREIMS [M+] m/z:628.3046; Calculated value about C38H44O8: 628.3036.

1H-NMR(600MHz，CDCl3)：δ12.55(1H，s，OH-6)，7.49(1H，d，J＝6.9Hz，H-10)，5.87(1H，t，J＝7.3Hz，H-27)，5.00(1H，br t，J＝6.3Hz，H-32)，3.46(1H，t，J＝5.6Hz，H-11)，3.27(1H，dd，J＝14.3，8.6Hz，H1-31)，3.17(1H，dd，J＝15.9，8.4Hz，H1-26)，3.10(1H，br dd，J＝13.9，4.4Hz，H2-31)，2.93(1H，m，H-3)，2.92(1H，m，H2-26)，2.49(1H，d，J＝9.3Hz，H-22)，2.40(1H，t，J＝8.4Hz，H-4)，2.30(1H，m，H-37)，2.28(1H，m，H1-21)，1.72(1H，m，H1-20)，1.70(3H，s，H-34)，1.68(3H，s，H-25)，1.66(3H，s，H-29)，1.60(3H，s，H-35)，1.58(1H，m，H1-36)，1.52(1H，m，H2-20)，1.50(1H，m，H2-36)，1.35(1H，dd，J＝13.3，9.6Hz，H2-21)，1.28(3H，s，H-19)，1.27(6H，s，H-24，H-40)，0.69(3H，s，H-39)。

13C-NMR(150MHz，CDCl3)：δ203.75(C-12)，178.86(C-8)，171.59(C-30)，161.51(C-18)，161.15(C-6)，155.01(C-16)，138.12(C-27)，134.58(C-10)，133.85(C-9)，130.79(C-33)，128.07(C-28)，122.35(C-32)，108.85(C-17)，105.28(C-5)，100.34(C-7)，90.32(C-14)，85.12(C-2)，84.20(C-23)，83.80(C-13)，48.93(C-22)，46.79(C-11)，46.19(C-37)，38.77(C-38)，38.53(C-20)，36.90(C-4)，34.97(C-3)，33.41(C-40)，29.90(C-25)，29.19(C-26)，28.91(C-24)，27.31(C-19)，25.73(C-35)，25.59(C-36)，25.29(C-21)，21.80(C-31)，20.77(C-29)，18.16(C-34)，17.65(C-39)。

The EIMS data presentation of product Gh-2607-1A: have a part peak [M]+position at m/z 628, and HREIMS data presentation [M]+m/z 628.3046, this represents that the molecular formula of product Gh-2607-1A is identical with above-mentioned gambogic acid, gambogellic acid and their epimer institute tool person, is all C38H44O8.

1H-NMR and the 13C-NMR spectrum display of product Gh-2607-1A, does not have cis two to replace double bond, does not have the terminal double link that pseudoallyl is rolled into a ball yet in the piperazine of product Gh-2607-1A mutters ring.

In addition, undertaken confirming rear discovery by 1H-1H COSY, HMQC and HMBC spectrum, product Gh-2607-1A is except monoterpene part, and other parts are all similar to gambogellic acid institute tool person.With gambogellic acid in comparison, product Gh-2607-1A has lacked a double bond, thus infers a product Gh-2607-1A structurally ring more than gambogellic acid.

The 1H-1H COSY spectroscopic data display of product Gh-2607-1A: 1 methine protons [δ 2.93 (1H, m, H-3)] is coupled with another methine protons [δ 2.40 (1H, t, J=8.4Hz, H-4)]; δ 2.40 and another methine protons [δ 2.30 (1H, m, H-37)] are coupled; δ 2.30 is coupled with 1 methene proton [δ 1.50 (1H, m, H2-36)] of C-36; Another methene proton [δ 1.72 (1H, m, H1-20)] is coupled with another methene proton [δ 1.58 (1H, m, H1-36)] of C-36.This proves that the monoterpene part of product Gh-2607-1A has 2 adjacent methylene groups [C-20 (δ 38.53) and C-36 (δ 25.59)] and 3 methin groups [C-37 (δ 46.19), C-4 (δ 36.90) and C-3 (δ 34.97)].The methin groups (C-3) of product Gh-2607-1A is the conversion of the methylene group (C-3) of transposition morellic acid, this conversion is form bond by C-3 and C-38 and instead of a terminal double link (terminal double bond) through inferring, and then form the structure of pinane type monoterpene, this structure contains 1 tetramethylene (cyclobutane), 1 together with-dimethyl group [C-39 (δ 17.65) and C-40 (δ 33.41), 1 contains oxygen level Four carbon [C-2 (δ 85.12)] and is affixed to this three grades of methyl groups [C-19 (δ 27.31)] containing oxygen level Four carbon.

Find from the HMBC spectroscopic data of product Gh-2607-1A: δ 2.93 (H-3), except being associated with aromatic nucleus carbon δ 105.28 (C-5), δ 161.15 (C-6) and δ 161.51 (C-18), is also associated with δ 85.12 (C-2), δ 46.19 (C-37), δ 38.77 (C-38), δ 38.53 (C-20), δ 36.90 (C-4), δ 33.41 (C-40) and δ 17.65 (C-39); δ 2.40 (H-4) is associated with δ 105.28 (C-5), δ 38.77 (C-38), δ 34.97 (C-3), δ 46.19 (C-37), δ 25.59 (C-36), δ 38.53 (C-20) and δ 27.31 (C-19); δ 2.30 (H-37) is associated with δ 105.28 (C-5), δ 85.12 (C-2), δ 38.77 (C-38), δ 34.97 (C-3), δ 36.90 (C-4), δ 38.53 (C-20), δ 25.59 (C-36) and δ 33.41 (C-40).HMBC spectroscopic data from above: the structure of the pinane type monoterpene that product Gh-2607-1A has links with the C-18 of ehter bond and aromatic nucleus in C-2 position, and C-4 and C-5 links, and the methyl group (C-19) be connected on C-2 position be equator to.

Find from the NOESY spectroscopic data of product Gh-2607-1A: δ 7.49 (H-10) is associated with δ 3.46 (H-11); δ 3.46 (H-11) is associated with δ 2.28 (H1-21); δ 1.35 (H2-21) is associated with δ 2.49 (H-22); δ 2.49 (H-22) is associated with δ 1.68 (H-25); δ 5.87 (H-27) is associated with δ 1.66 (H-29).This proves that the three-dimensional arrangement of this part is 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are for trans, and double bond Δ 27,28 is Z configuration.In addition, δ 2.93 (H-3) is associated with δ 2.40 (H-4), δ 1.28 (H-19) and δ 1.27 (H-40); δ 2.40 (H-4) is associated with δ 2.30 (H-37), δ 1.27 (H-40) and δ 1.66 (H-29); δ 2.30 (H-37) is associated with δ 1.66 (H-29) and δ 1.70 (H-34); δ 1.50 (H2-36) is associated with δ 1.66 (H-29).It can thus be appreciated that C-2 is S configuration, and in pinane structure, 3 methine protons H-3, H-4 and H-37 be positioned on tetramethylene ring are all cis (cis), and its three-dimensional arrangement is 3R, 4R, 37S configuration.In addition, also find from NOESY spectroscopic data: δ 3.27 (H1-31) and δ 3.10 (H2-31) is all associated with δ 1.66 (H-29); δ 3.17 (H1-26) and δ 2.92 (H2-26) is all associated with δ 1.70 (H-34); δ 0.69 (H-39) is associated with δ 1.72 (H1-20), δ 1.28 (H-19) and δ 1.27 (H-40).

Comprehensive above data, product Gh-2607-1A is identified is a compounds with the following chemical structure formula:

Product Gh-2607-1A is named as epiformoxanthone B (epiformoxanthoneB), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10R, 12S, 15S, 18aS)-3a, 4, 5, 7, 12, 13, 14, 15-octahydro-8-hydroxyl-3, 3, 11, 11, 15-pentamethyl--17-(3-methyl-2-butene base)-7, 19-dioxy-1, 5-methylene radical-1H, 3H, 9H-furo [3.4-g]-16-oxygen-ginseng ring [4.4.0.09, 12] decyl also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10R, 12S, 15S, 18aS)-3a, 4, 5, 7, 12, 13, 14, 15-octahydro-8-hydroxy-3, 3, 11, 11, 15-pentamethyl-17-(3-methyl-2-butenyl)-7, 19-dioxo-1, 5-methano-1H, 3H, 9H-furo [3.4-g]-16-oxa-tricyclo [4.4.0.09, 12] decano [3.2-b] xanthen-1-yl]-, (2Z)-].

15. product Gh-2508:

Product Gh-2508 is as follows by the character recorded:

Yellow powder, fusing point: 113-118 DEG C.

EIMS m/z (relative intensity): 628 [M]+(71), 600 (100), 575 (14), 545 (52), 517 (68), 501 (12), 473 (96), 459 (12), 431 (26), 417 (18), 389 (29), 355 (64), 349 (26), 307 (12), 295 (22), 253 (24), 245 (29), 214.9 (26), 189 (17), 105 (15), 91 (18), 69 (35).

1H-NMR(600MHz，CDCl3)：δ12.57(1H，s，OH-6)，7.50(1H，d，J＝6.9Hz，H-10)，5.79(1H，dd，J＝16.0，7.2Hz，H-27)，5.30(1H，d，J＝16.0Hz，H-26)，5.14(1H，br t，J＝6.9Hz，H-32)，4.57(1H，s，H1-40)，4.23(1H，s，H2-40)，3.50(1H，br d，J＝2.8Hz，H-4)，3.43(1H，dd，J＝6.8，4.4Hz，H-11)，3.23(2H，m，H-31)，2.90(1H，dq，J＝7.2，7.1Hz，H-28)，2.56(1H，d，J＝9.3Hz，H-22)，2.29(1H，dd，J＝13.4，4.6Hz，H1-21)， 2.16(1H，br d，J＝12.5Hz，H-37)，1.94(1H，br d，J＝12.9Hz，H1-20)，1.89(1H，m，H1-3)，1.86(3H，s，H-39)，1.75(1H，m，H2-3)，1.73(6H，s，H-25，H-34)，1.65(3H，s，H-35)，1.56(1H，dt，J＝13.4，5.0Hz，H2-20)，1.42(1H，m，H2-21)，1.38(3H，s，H-19)，1.36(2H，m，H-36)，1.27(3H，s，H-24)，0.90(3H，d，J＝7.1Hz，H-29)。

13C-NMR(150MHz，CDCl3)：δ203.38(C-12)，179.21(C-8)，177.35(C-30)，164.29(C-18)，160.61(C-6)，155.79(C-16)，147.66(C-38)，134.54(C-9，C-10)，134.33(C-27)，131.17(C-33)，123.80(C-26)，122.45(C-32)，108.83(C-40)，106.65(C-17)，104.18(C-5)，100.11(C-7)，91.46(C-14)，84.76(C-13)，84.13(C-23)，77.07(C-2)，48.27(C-22)，48.13(C-37)，47.06(C-11)，41.74(C-28)，39.31(C-20)，36.92(C-3)，30.03(C-25)，28.94(C-4)，28.73(C-24)，28.50(C-19)，25.74(C-35)，25.51(C-21)，22.98(C-39)，22.74(C-36)，21.94(C-31)，18.18(C-34)，15.80(C-29)。

The EIMS data presentation of product Gh-2508: have a part peak [M]+position at m/z 628, cataclasm graphic (fragmentationpatterns) of this and product Gh-2603-2 (namely, gambogellic acid) is similar.In addition, 1H-NMR and the 13C-NMR spectrum of product Gh-2508 is similar to product Gh-2603-2 institute tool person haply.

1H-NMR and the 13C-NMR spectrum display of product Gh-2508: the side chain containing carboxylic group be connected on C-13 position is different from product Gh-2603-2 institute tool person, two replace double bond Δ 26,27 [δ 5.79 (1H, dd, J=16.0,7.2Hz, H-27), δ 134.33 (C-27) and δ 5.30 (1H, d, J=16.0Hz, H-26), δ 123.80 (C-26)] instead of three of product Gh-2603-2 and replace double bond (trisubstituted double bond) Δ 27,28 and methylene groups (C-26).In addition, the coupling constant (J) of 2 thiazolinyl protons (olefinic protons) (H-26 and H-27) be coupled mutually is 16.0Hz, so Δ 26,27 is trans double bond (namely, E configuration).Known by the doublet signal of δ 0.90 (3H, d, J=7.1Hz, H-29): C-29 changes secondary methyl group into, H-29 and methine protons [δ 2.90 (1H, dq, J=7.2,7.1Hz, H-28)] are coupled.In addition, methin groups [δ 2.90 (H-28), δ 41.74 (C-28)] adjacent with carboxyl carbonyl (carboxyl carbon) [δ 177.35 (C-30)], this represents that carboxyl carbon has not been α, beta-unsaturated carbonyl carbon (α, β-unsaturated carbonyl carbon).

Find from the HMBC spectroscopic data of product Gh-2508: δ 5.79 (H-27) is associated with δ 177.35 (C-30), δ 15.80 (C-29), δ 41.74 (C-28) and δ 84.76 (C-13); δ 5.30 (H-26) is associated with δ 177.35 (C-30), δ 41.74 (C-28), δ 134.33 (C-27) and δ 84.76 (C-13); δ 2.90 (H-28) is associated with δ 177.35 (C-30), δ 15.80 (C-29), δ 134.33 (C-27) and δ 123.80 (C-26); δ 0.90 (H-29) is associated with δ 177.35 (C-30) and δ 134.33 (C-27).This proves that the side chain of C-13 is (E)-2-methyl-3-butenoic acid [(E)-2-methyl-3-butenoic acid].

Find from the NOESY spectroscopic data of product Gh-2508: δ 7.50 (H-10) is associated with δ 3.43 (H-11); δ 3.43 (H-11) is associated with δ 2.29 (H1-21); δ 2.56 (H-22) is associated with δ 1.42 (H2-21) and δ 1.27 (H-24).This proves that the three-dimensional arrangement of this part is identical with gambogic acid or gambogellic acid institute tool person, is all 11S, 13R, 14S, 22S configuration.In addition, δ 0.90 (H-29), except being associated with δ 5.79 (H-27) and δ 5.30 (H-26), is also associated with adjacent δ 2.90 (H-28).

Undertaken confirming rear discovery by 1H-1H COSY, HMQC and HMBC spectrum, the monoterpene part of product Gh-2508 is identical with product Gh-2603-2 institute tool person.Be affixed to the methine protons [δ 2.16 (1H of lsopropenyl group, br d, J=12.5Hz, H-37) signal display], the methine protons and the neighbour proton (vicinal proton) that are affixed to lsopropenyl group have sizable neighbour coupling constant (vicinal coupling constant) (J=12.5Hz), they are couplings of axial-axial (axial-axial), so H-37 is axially, and S configuration.

In addition, also find from NOESY spectroscopic data: δ 3.50 (H-4), except being associated with δ 2.16 (H-37), is also associated with δ 1.89 (H1-3) and δ 1.75 (H2-3); δ 1.89 (H1-3) is associated with δ 1.38 (H-19); Axial hydrogen δ 2.16 (H-37) is associated with δ 1.89 (H1-3) and δ 1.56 (H2-20).This represents that monoterpene ring is 1,3-bis-axial (1, chair form configuration (chair conformation) 3-diaxial), and the methyl proton (H-19) be connected on C-2 position and the lsopropenyl group be connected on C-37 position be all equator to.In addition, due to the methyl proton [δ 0.90 (H-29)] Yu monoterpene part (monoterpene moiety) that are connected to the side chain on C-13 position lsopropenyl group outer-methylene radical (exo-methylene) proton [δ 4.57 (H1-40) and δ 4.23 (H2-40)] do not have core Ao Fohaose effect (nuclear Overhauser effect, NOE), thus estimating C-2 is R configuration.This proves that the three-dimensional arrangement of this monoterpene part is 2R, 4R, 37S.

Comprehensive above data, product Gh-2508 is identified is a compounds with the following chemical structure formula:

Product Gh-2508 is named as β-gambogellic acid (β-gambogellic acid), and { IUPAC names: 3-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10S, 13R, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxyl-3, 3, 13-trimethylammonium-15-(3-methyl-2-butene base)-10-(1-methyl ethylene)-7, 18-dioxy-1, 5:9, 13-dimethylene-1H, 3H, 9H-furo [3.4-g] oxygen is [3.2-b]-1-base also]-, (3E)-[3-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10S, 13R, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxy-3, 3, 13-trimethyl-15-(3-methyl-2-butenyl)-10-(1-methylethenyl)-7, 18-dioxo-1, 5:9, 13-dimethano-1H, 3H, 9H-furo [3.4-g] oxocino [3.2-b] xanthen-1-yl]-, (3E)-].

16. product Gh-2507:

Product Gh-2507 is as follows by the character recorded:

Yellow needles, fusing point: 148-152 DEG C.

EIMS m/z (relative intensity): 628 [M]+(48), 600 (100), 585 (9), 545 (41), 517 (69), 510 (12), 473 (93), 431 (24), 417 (18), 389 (28), 355 (68), 347 (25), 307 (11), 299 (24), 295 (20), 253 (22), 245 (28), 214.9 (22), 199 (18), 189 (15), 105 (12), 91 (17), 69 (25).

1H-NMR(600MHz，CDCl3)：δ12.61(1H，s，OH-6)，7.48(1H，d，J＝6.9Hz，H-10)，6.06(1H，dd，J＝15.7，7.2Hz，H-27)，5.15(1H，d，J＝15.7Hz，H-26)，5.11(1H，br t，J＝6.9Hz，H-32)，4.56(1H，s，H1-40)，4.20(1H，s，H2-40)，3.54(1H，br d，J＝2.7Hz，H-4)，3.44(1H，dd，J＝6.8，4.5Hz，H-11)，3.25(1H，dd，J＝14.4，8.1Hz，H1-31)，3.17(1H，dd，J＝14.4，5.6Hz，H2-31)，2.90(1H，dq，J＝7.2，7.1Hz，H-28)，2.56(1H，d，J＝9.3Hz，H-22)，2.30(1H，dd，J＝13.5，4.7Hz，H1-21)，2.16(1H，br d，J＝12.4Hz，H-37)，1.97(1H，br d，J＝12.8Hz，H1-20)，1.88(1H，m，H1-3)，1.87(3H，s，H-39)，1.74(1H，m，H2-3)，1.72(3H，s，H-25)，1.71(3H，s，H-34)，1.64(3H，s，H-35)，1.55(1H，d t，J＝13.5，4.9Hz，H2-20)，1.44(1H，m，H2-21)，1.43(1H，m，H1-36)，1.36(3H，s，H-19)，1.32(1H，m，H2-36)，1.27(3H，s，H-24)，0.92(3H，d，J＝7.0Hz，H-29)。

13C-NMR(150MHz，CDC l 3)：δ203.53(C-12)，179.16(C-8)，177.27(C-30)，164.37(C-18)，160.73(C-6)，156.10(C-16)，148.04(C-38)，134.80(C-10)，134.65(C-27)，134.52(C-9)，131.20(C-33)，122.91(C-26)，122.39(C-32)，108.71(C-40)，106.53(C-17)，104.14(C-5)，99.93(C-7)，90.92(C-14)，84.66(C-13)，84.10(C-23)，77.16(C-2)，48.49(C-22)，48.15(C-37)，47.03(C-11)，42.00(C-28)，39.21(C-20)，37.04(C-3)，29.96(C-25)，28.93(C-4)，28.76(C-24)，28.53(C-19)，25.77(C-35)，25.35(C-21)，22.94(C-39)，22.59(C-36)，22.01(C-31)，18.15(C-34)，15.92(C-29)。

The EIMS data presentation of product Gh-2507: have a part peak [M]+position at m/z 628, this and product Gh-2603-1 are (namely, epigambogellic acid) and Gh-2603-2 (namely, gambogellic acid) institute tool person identical.In addition, 1H-NMR and the 13C-NMR spectrum of product Gh-2507 is similar to product Gh-2603-1 institute tool person haply.

1H-NMR and the 13C-NMR spectrum display of product Gh-2507: the side chain containing carboxylic group be connected on C-13 position is different from product Gh-2603-1 institute tool person, two replace double bond Δ 26,27 [δ 5.15 (1H, d, J=15.7Hz, H-26), δ 122.91 (C-26) and δ 6.06 (1H, dd, J=15.7,7.2Hz, H-27), δ 134.65 (C-27)] instead of three of product Gh-2603-1 and replace double bond Δ 27,28 and methylene groups (C-26).The coupling constant (J) of 2 thiazolinyl protons (H-26 and H-27) be coupled mutually is 15.7Hz, so Δ 26,27 is trans double bond (namely, E configuration).By δ 0.92 (3H, d, J=7.0Hz, H-29) doublet signal is known: C-29 changes secondary methyl group into, proton (H-29) and methine protons [δ 2.90 (1H, the dq of this secondary methyl group, J=7.2,7.1Hz, H-28)] coupling.In addition, because methin groups [δ 2.90 (H-28), δ 42.00 (C-28)] is adjacent with carboxyl carbon [δ 177.27 (C-30)], this represents that carboxyl carbon has not been α, beta-unsaturated carbonyl carbon.

Find from the HMBC spectroscopic data of product Gh-2507: δ 6.06 (H-27) is associated with δ 177.27 (C-30), δ 15.92 (C-29), δ 42.00 (C-28) and δ 84.66 (C-13); δ 5.15 (H-26) is associated with δ 42.00 (C-28), δ 134.65 (C-27), δ 84.66 (C-13) and δ 203.53 (C-12); δ 2.90 (H-28) is associated with δ 177.27 (C-30), δ 15.92 (C-29), δ 134.65 (C-27) and δ 122.91 (C-26); δ 0.92 (H-29) is associated with δ 177.27 (C-30), δ 42.00 (C-28) and δ 134.65 (C-27).This proves that the side chain of C-13 is (E)-2-methyl-3-butenoic acid [(E)-2-methyl-3-butenoic acid].

Find from the NOESY spectroscopic data of product Gh-2507: δ 7.48 (H-10) is associated with δ 3.44 (H-11); δ 3.44 (H-11) is associated with δ 2.30 (H1-21) and δ 1.44 (H2-21); δ 1.44 (H2-21) is associated with δ 2.56 (H-22); δ 2.56 (H-22) is associated with δ 1.72 (H-25).This proves that the three-dimensional arrangement of this part is identical with gambogic acid, gambogellic acid and epigambogellic acid institute tool person, is all 11S, 13R, 14S, 22S configuration.In addition, δ 0.92 (H-29) is associated with δ 2.90 (H-28), δ 6.06 (H-27), δ 5.15 (H-26) and δ 4.20 (H2-40).The lsopropenyl group of methyl proton (H-29) and monoterpene part outer-methene proton [δ 4.56 (H1-40), δ 4.20 (H2-40)] is associated, thus estimating C-2 is S configuration, if C-2 is R configuration, there will not be intersection peak (cross peak).

Undertaken confirming rear discovery by 1H-1H COSY, HMQC and HMBC spectrum, the monoterpene part of product Gh-2507 is identical with product Gh-2603-1 institute tool person.Be affixed to the methine protons [δ 2.16 (1H of lsopropenyl group, br d, J=12.4Hz, H-37) signal display], the methine protons and the neighbour proton (vicinal proton) that are affixed to lsopropenyl group have sizable neighbour coupling constant (vicinal coupling constant) (J=12.4Hz), they are couplings of axial-axial (axial-axial), so H-37 is axially, and R configuration.

In addition, NOESY spectroscopic data also finds: δ 3.54 (H-4), except being associated with δ 2.16 (H-37), is also associated with δ 1.88 (H1-3) and δ 1.74 (H2-3); And methene proton (H1-3 and H2-3) is all associated with δ 1.36 (H-19); Axial hydrogen δ 2.16 (H-37), except being associated with δ 1.87 (H-39), is also associated with δ 1.88 (H1-3), δ 1.55 (H2-20) and δ 1.43 (H1-36).This proves that monoterpene ring is 1, the chair form configuration of 3-bis-axis, and the methyl group (H-19) be connected on C-2 position and the lsopropenyl group be connected on C-37 position be all equator to, therefore the three-dimensional arrangement of p-ene-type in Meng monoterpene is 2S, 4S, 37R configuration.

Comprehensive above data, product Gh-2507 is identified is a compounds with the following chemical structure formula:

Product Gh-2507 is named as β-epigambogellic acid (β-epigambogellicacid), and { IUPAC names: 3-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10R, 13S, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxyl-3, 3, 13-trimethylammonium-15-(3-methyl-2-butene base)-10-(1-methyl ethylene)-7, 18-dioxy-1, 5:9, 13-dimethylene-1H, 3H, 9H-furo [3.4-g] oxygen is [3.2-b]-1-base also]-, (3E)-[3-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9S, 10R, 13S, 16aS)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8-hydroxy-3, 3, 13-trimethyl-15-(3-methyl-2-butenyl)-10-(1-methylethenyl)-7, 18-dioxo-1, 5:9, 13-dimethano-1H, 3H, 9H-furo [3.4-g] oxocino [3.2-b] xanthen-1-yl]-, (3E)-].

17. product Gh-2501:

Product Gh-2501 is as follows by the character recorded:

Yellow powder, fusing point: 100-103 DEG C.

EIMS m/z (relative intensity): 544 [M]+(100), 529 (67), 516 (16), 501 (56), 473 (13), 435 (11), 405 (59), 389 (25), 363 (43), 349 (37), 337 (12), 307 (34), 287 (58), 259 (29), 229 (69), 215 (59), 189 (35), 147 (23), 135 (30), 105 (42), 91 (27), 83 (29), 69 (36), 55 (34).

1H-NMR(600MHz，CDCl3)：δ12.70(1H，s)，9.21(1H，s)，7.54(1H，d，J＝7.0Hz)，6.59(1H，d，J＝10.0Hz)，6.37(1H，t，J＝8.5Hz)，5.50(1H，d，J＝10.0Hz)，5.07(1H，t，J＝8.4Hz)，3.51(1H，dd，J＝6.9，4.5Hz)，3.25(1H，dd，J＝14.4，8.1Hz)，3.17(1H，br dd，J＝13.9，5.9Hz)， 2.71(1H，ddd，J＝16.0，7.5，0.8Hz)，2.62(1H，ddd，J＝16.0，7.0，0.9Hz)，2.56(1H，d，J＝9.4Hz)，2.34(1H，dd，J＝13.6，4.7Hz)，1.73(3H，s)，1.71(3H，s)，1.62(3H，s)，1.43(3H，s)，1.41(3H，s)，1.39(1H，m)，1.29(3H，s)，1.28(3H，d，J＝1.2Hz)。

13C-NMR(150MHz，CDCl3)：δ203.02，194.46，178.84，161.34，157.68，157.15，146.51，140.11，135.62，133.34，131.98，126.39，121.82，115.28，108.07，103.27，100.36，90.79，83.98，83.39，78.87，48.99，46.85，29.96，28.96，28.93，28.39(2C)，25.75，25.26，21.67，18.16，8.58。

According to measured spectroscopic data, product Gh-2501 is identified is a known compound with the following chemical structure formula, namely isomorellin (isomorellin):

18. product Gh-2505:

Product Gh-2505 is as follows by the character recorded:

Yellow powder, fusing point: 67-70 DEG C.

EIMS m/z (relative intensity): 630 [M]+(100), 602 (19), 545 (14), 507 (36), 479 (22), 475 (18), 433 (8), 351 (27), 309 (17), 295 (38), 253 (45), 245 (16), 231 (16), 213 (13), 177 (15), 147 (8), 69 (29).

1H-NMR(600MHz，CDCl3)：δ12.81(1H，s)，7.52(1H，d，J＝6.9Hz)，6.48(1H，s)，5.83(1H，t，J＝7.2Hz)，5.17(1H，t，J＝6.9Hz)，5.06(1H，br t，J＝6.6Hz)，5.02(1H br t，J＝6.7Hz)，3.48(1H，dd，J＝6.6，4.8Hz)，3.30(1H，m)，3.28(2H，m)，3.24(1H，dd，J＝16.1，7.1Hz)，3.10(1H，dd，J＝15.7，8.7Hz)，2.86(1H，ddd，J＝15.9，6.7，1.1Hz)，2.49(1H，d，J＝9.4Hz)，2.30(1H，dd，J＝13.5，4.7Hz)，2.06(2H，m)，2.00(2H，m)，1.74(3H，s)，1.71(3H，s)，1.70(3H，s)，1.66(3H，s)，1.65(3H，s)，1.64(3H，s)，1.56(3H，s)，1.36(1H，dd，J＝13.5，9.5Hz)，1.27(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.38，179.12，170.39，163.62，160.34，155.85，139.06，136.95，135.15，133.91，133.52，131.88，128.50，123.83，121.91，121.35，107.50，106.43，100.64，90.41，83.94，83.87，48.90，46.88，39.68，29.79，29.46，28.92，26.33，25.71， 25.67，25.21，22.03，21.09，20.74，17.98，17.68，16.16。

According to measured spectroscopic data, product Gh-2505 is identified is a known compound with the following chemical structure formula, namely gambogenic acid (gambogenic acid):

19. product Gh-2642:

Product Gh-2642 is as follows by the character recorded:

Yellow powder, fusing point: 157-159 DEG C.

EIMS m/z (relative intensity): 630 [M]+(100), 602 (11), 545 (11), 533 (16), 507 (46), 479 (21), 475 (15), 433 (7), 419 (8), 381 (9), 357 (13), 351 (20), 309 (14), 295 (27), 253 (34), 245 (15), 231 (16), 213 (11), 177 (15), 147 (8), 135 (8), 105 (11), 69 (44).

1H-NMR(600MHz，CDCl3)：δ12.76(1H，s)，7.52(1H，d，J＝7.0Hz)，6.70(1H，br s)，6.59(1H，t，J＝7.5Hz)，5.19(1H，t，J＝7.0Hz)，5.10(1H，br t，J＝6.0Hz)，5.02(1H，br t，J＝6.1Hz)，3.47(1H，t，J＝5.6Hz)，3.34(2H，m)，3.31(2H，m)，2.60(1H，dd，J＝15.7，7.9Hz)，2.50(1H，m)，2.49(1H，d，J＝9.5Hz)，2.30(1H，dd，J＝13.3，4.4Hz)，2.06(2H，m)，2.02(2H，m)，1.76(3H，s)，1.70(3H，s)，1.66(6H，s)，1.62(3H，s)，1.54(3H，s)，1.35(1H，dd，J＝13.3，9.5Hz)，1.30(3H，s)，1.25(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.15，179.02，172.20，163.51，160.27，155.87，138.98，136.90，135.50，133.72，133.33，131.75，128.64，123.81，121.88，121.15，107.51，106.63，100.55，90.37，83.65，83.49，48.89，46.82，39.61，29.80，28.85(2C)，26.27，25.65，25.60，25.19，21.98，21.10，17.93，17.59，16.16，11.34。

According to measured spectroscopic data, product Gh-2642 is identified is a known compound with the following chemical structure formula, namely isogambogenic acid (isogambogenic acid):

20. product Gh-1601-A:

Product Gh-1601-A is as follows by the character recorded:

Yellow powder, fusing point: 143-145 DEG C.

EIMS m/z (relative intensity): 644 [M]+(72), 598 (18), 561 (100), 515 (23), 474 (35), 431 (9), 391 (10), 355 (14), 349 (7), 347 (6), 253 (6), 248 (11), 215 (11), 189 (6), 125 (4), 69 (18).

1H-NMR(600MHz，CDCl3)：δ12.76(1H，s)，7.54(1H，d，J＝6.9Hz)，6.61(1H，d，J＝10.2Hz)，6.31(1H，t，J＝7.2Hz)，5.41(1H，d，J＝10.1Hz)，5.02(2H，br s)，4.09(1H，d，J＝13.3Hz)，4.01(1H，d，J＝13.3Hz)，3.49(1H，br t，J＝5.6Hz)，3.27(1H，dd，J＝14.2，8.0Hz)，3.13(1H，br dd，J＝14.3，5.2Hz)，2.96(2H，d，J＝7.5Hz)，2.51(1H，d，J＝9.6Hz)，2.31(1H，dd，J＝13.4，4.7Hz)，2.00(2H，m)，1.72(1H，m)，1.70(3H，s)，1.67(3H，s)，1.62(3H，s)，1.60(3H，s)，1.58(1H，m)，1.52(3H，s)，1.38(1H，m)，1.37(3H，s)，1.26(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.04，179.01，169.43，161.66，157.50，157.27，139.33，135.66，133.20，131.90，131.87，131.24，124.80，123.69，121.96，115.76，107.75，102.82，100.47，90.76，84.25，83.67，81.47，64.80，48.92，46.82，41.93，29.82，29.14，28.82，27.76，25.68，25.65，25.12，22.70，21.60，18.13，17.61。

According to measured spectroscopic data, product Gh-1601-A is identified is a known compound with the following chemical structure formula, namely 30-hydroxygambogic acid (30-hydroxygambogic acid):

21. product Gh-1602:

Product Gh-1602 is as follows by the character recorded:

Yellow powder, fusing point: 98-100 DEG C.

EIMS m/z (relative intensity): 644 [M]+(28), 598 (5), 561 (100), 515 (15), 474 (9), 431 (3), 389 (5), 355 (5), 347 (5), 253 (3), 245 (6), 215 (10), 189 (5), 125 (3), 69 (8).

1H-NMR(600MHz，CDCl3)：δ12.73(1H，s)，7.54(1H，d，J＝6.9Hz)，6.60(1H，d，J＝10.1Hz)，6.40(1H，t，J＝7.4Hz)，5.39(1H，d，J＝10.2Hz)，5.07(1H，t，J＝7.0Hz)，5.01(1H，br t，J＝6.8Hz)，4.09(1H， d，J＝13.1Hz)，4.02(1H，d，J＝13.2Hz)，3.46(1H，t，J＝5.6Hz)，3.27(1H，dd，J＝14.6，8.2Hz)，3.12(1H，dd，J＝14.6，5.0Hz)，2.96(2H，d，J＝7.5Hz)，2.50(1H，d，J＝9.3Hz)，2.29(1H，dd，J＝13.5，4.7Hz)，2.04(2H，m)，1.73(1H，m)，1.71(3H，s)，1.67(3H，s)，1.64(3H，s)，1.62(1H，m)，1.61(3H，s)，1.56(3H，s)，1.36(1H，m)，1.32(3H，s)，1.26(3H，s)。

13C-NMR(150MHz，CDCl3)：δ203.08，179.02，169.57，161.46，157.50，157.24，140.20，135.91，133.02，132.20，131.62，131.15，124.94，123.74，122.04，115.81，107.89，102.95，100.50，90.89，84.02，83.56，81.21，64.64，48.91，46.82，41.67，29.90，29.14，28.77，26.91，25.69，25.61，25.19，22.73，21.58，18.15，17.60。

According to measured spectroscopic data, product Gh-1602 is identified is a known compound with the following chemical structure formula, namely 30-hydroxyepigambogic acid (30-hydroxyepigambogicacid):

22. product Gh-2641-1:

Product Gh-2641-1 is as follows by the character recorded:

Yellow powder, fusing point: 94-98 DEG C.

EIMS m/z (relative intensity): 646 [M]+(100), 545 (18), 523 (80), 495 (28), 477 (17), 449 (16), 367 (44), 349 (20), 325 (37), 295 (41), 252.9 (49), 245 (27), 213 (32), 147 (16).

1H-NMR(600MHz，CDCl3)：δ12.84(1H，s，OH-6)，7.46(1H，d，J＝7.0Hz，H-10)，5.30(1H，ddd，J＝12.1，4.0，1.4Hz，H-27)，5.19(1H，dt，J＝7.1，1.2Hz，H-32)，5.04(1H，tt，J＝6.9，1.3Hz，H-37)，4.68(1H，dd，J＝9.8，6.5Hz，H-4)，3.76(1H，dd，J＝14.5，12.1Hz，H1-26)，3.48(1H，d，J＝7.0，4.2Hz，H-11)，3.16(2H，m，H-31)，3.13(1H，dd，J＝15.3，6.5Hz，H1-3)，3.03(1H，dd，J＝15.3，9.8Hz，H2-3)，2.76(1H，ddd，J＝14.5，4.0，2.2Hz，H2-26)，2.39(1H，d，J＝9.6Hz，H-22)，2.32(1H，dd，J＝13.5，4.5Hz，H1-21)，2.02(2H，m，H-36)，1.91(2H，m，H-20)，1.70(3H，s，H-35)，1.64(3H，s，H-25)，1.61(3H，s，H-39)，1.53(3H，s，H-40)，1.51(3H，s，H-29)，1.43(3H，s，H-19)，1.33(1H，m，H2-21)， 1.28(3H，s，H-24)，1.25(3H，s，H-34)。

13C-NMR(150MHz，CDCl3)：δ202.10(C-12)，178.09(C-8)，168.66(C-30)，167.88(C-18)，163.12(C-16)，152.96(C-6)，135.41(C-33)，135.12(C-27)，134.13(C-10)，133.72(C-9)，131.18(C-38)，129.76(C-28)，124.36(C-37)，121.53(C-32)，104.85(C-5)，103.47(C-17)，100.53(C-7)，90.43(C-4)，90.21(C-14)，84.37(C-13)，83.66(C-23)，73.26(C-2)，48.88(C-22)，46.61(C-11)，39.67(C-20)，30.13(C-26)，29.92(C-25)，29.15(C-24)，26.65(C-36，C-3)，26.60(C-19)，25.60(C-39)，25.19(C-21)，24.65(C-35)，21.25(C-31)，20.47(C-29)，17.58(C-40)，16.04(C-34)。

The EIMS data presentation of product Gh-2641-1: have a part peak [M]+position at m/z 646, this is equivalent to the yellow pigment (xanthones) that a part formula is C38H46O9.

The 1H-NMR spectrum display of product Gh-2641-1: it has 1 chelated hydroxy group (chelated hydroxy group) (δ 12.84), 4 thiazolinyl protons (olefinic protons) (δ 7.46, δ 5.30, δ 5.19 and δ 5.04) and 1 primary hydroxyl groups (δ 4.68).With known gambogic acid in comparison, the 1H-NMR spectrum of product Gh-2641-1 has lacked 1 cis-two be coupled mutually and has replaced double bond (cis-disubstituted double bond) signal, and many 1 AX2 spin system (AX2 spin system) proton signal [δ 4.68 (1H, dd, J=9.8, 6.5Hz, H-4), δ 3.13 (1H, dd, J=15.3, 6.5Hz, and δ 3.03 (1H H1-3), dd, J=15.3, 9.8Hz, H2-3)], this display product Gh-2641-1 may be known neogambogic acid (neogambogic acid).

Except utilizing 1H-1H COSY spectrum to confirm except decouple protons in above-mentioned 1H-NMR spectrum relevant (coupled proton correlations), associating of proton and carbon signal can be obtained from HMQC spectrum.In addition, find from the HMBC spectroscopic data of product Gh-2641-1: hydroxyl methine protons (hydroxymethine proton) [δ 4.68 (1H, dd, J=9.8,6.5Hz, H-4)] except being associated with 2 level Four carbon [δ 103.47 (C-17) and δ 167.88 (C-18)], being associated containing oxygen level Four carbon [δ 73.26 (C-2)] and three grades of methyl groups (tertiary methylgroup) [δ 26.60 (C-19)] of ring of also muttering with piperazine.This proves that oh group is on C-4 position, and by the coupling constant (J=9.8 of H-4,6.5Hz) known, accurate-axially (quasi-axial) C-4 proton respectively with accurate-axial H2-3 (J=9.8Hz) and standard-equator to H1-3 (J=6.5Hz) be coupled, so oh group is β-orientation (β-orientation).

In addition, find from the NOESY spectroscopic data of product Gh-2641-1: δ 7.46 (H-10) is associated with δ 3.48 (H-11); δ 3.48 (H-11) is associated with δ 2.32 (H1-21); δ 2.32 (H1-21) is associated with δ 2.39 (H-22); δ 2.39 (H-22) is associated with δ 1.28 (H-24); δ 5.30 (H-27) is associated with δ 1.51 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are for trans, and double bond Δ 27,28 is Z configuration.In addition, δ 4.68 (H-4) is associated with δ 1.43 (H-19), and this represents that the methyl proton be connected on C-2 position is axially (α-), and identical with gambogic acid institute tool person, is all R configuration.

Comprehensive above data, product Gh-2641-1 is identified is a known compound with the following chemical structure formula, namely neogambogic acid (neogambogic acid):

23. product Gh-1631:

Product Gh-1631 is as follows by the character recorded:

Yellow needles, fusing point: 95-97 DEG C.

EIMS m/z (relative intensity): 646 [M]+(54), 618 (75), 573 (12), 545 (8), 520 (13), 492 (75), 491 (100), 477 (18), 449 (38), 373 (19), 349 (11), 321 (15), 295 (27), 267 (10), 252.9 (37), 245 (18), 213 (11), 188.9 (15), 176.9 (11), 109 (24), 99 (20), 69 (83).HREIMS [M]+m/z 646.3146; Calculated value about C38H46O9: 646.3142.

1H-NMR(600MHz，CDCl3)：δ12.45(1H，s，OH-6)，7.42(1H，d，J＝6.9Hz，H-10)，5.32(1H，br d，J＝9.9Hz，H-27)，5.18(1H，t，J＝6.8Hz，H-32)，5.08(1H，t，J＝7.1Hz，H-37)，4.75(1H，t，J＝8.1Hz，H-3)，3.53(1H，dd，J＝15.7，11.0Hz，H1-26)，3.48(1H，dd，J＝6.6，4.8Hz，H-11)，3.29(1H，dd，J＝15.1，7.3Hz，H1-31)，3.24(1H，dd，J＝15.2，6.3Hz，H2-31)，3.07(2H，d，J＝8.1Hz，H-4)，2.69(1H，ddd，J＝15.8，4.0，2.3Hz，H2-26)，2.53(1H，d，J＝9.4Hz，H-22)，2.30(1H，dd，J＝13.5，4.8Hz，H1-21)，2.09(1H，m，H1-36)，2.02(1H，m，H2-36)，1.72(3H，s，H-34)，1.67(3H，s，H-35)，1.66(3H，s，H-39)，1.65(3H，s，H-25)，1.601(3H，s，H-40)，1.597(3H，sh，H-29)，1.57(1H，m，H1-20)，1.47(1H，m，H2-20)，1.42(1H，m，H2-21)，1.42(3H，s，H-19)，1.23(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ204.15(C-12)，179.64(C-8)，168.08(C-30)，168.02(C-18)，158.45(C-16)，157.32(C-6)，136.74(C-27，C-33)，134.37(C-10)，132.40(C-38)，132.12(C-9)，128.40(C-28)，123.56(C-37)，121.83(C-32)，105.90(C-5)，103.96(C-17)，101.72(C-7)，90.28(C-14)，90.00(C-3)，84.40(C-13)，83.45(C-23)，75.77 (C-2)，75.70(C-2)，48.78(C-22)，47.71(C-11)，36.93(C-20)，29.87(C-25)，29.32(C-26)，28.87(C-24)，25.93(C-4)，25.71(C-35)，25.64(C-39)，25.14(C-21)，23.54(C-19)，22.51(C-31)，22.06(C-36)，20.91(C-29)，17.94(C-34)，17.66(C-40)。

The EIMS data presentation of product Gh-1631: have a part peak [M]+position in m/z 646 (54) and a benchmark peak position at m/z 491 (100); And HREIMS data presentation [M]+m/z 646.3146, tool person is identical for the molecular formula of this expression product Gh-1631 and product Gh-2641-1 (namely, neogambogic acid) institute, is all C38H46O9.

The 1H-NMR spectrum of product Gh-1631 shows it and has 1 chelated hydroxy group (chelatedhydroxy group) (δ 12.45), 4 thiazolinyl proton (olefinic protons) (δ 7.42, δ 5.32, δ 5.18, δ 5.08) and 1 there is hydroxyl methine protons [δ 4.75 (1H, t, J=8.1Hz)] with methene proton [δ 3.07 (2H, d, J=8.1Hz)] primary hydroxyl groups (secondary hydroxy group) that intercouples.The 1H-NMR spectrum of product Gh-1631 is similar to product Gh-2641-1 institute tool person haply.

Find from HMQC spectroscopic data: δ 90.00 (-OCH-) and δ 25.93 (-CH2-) is the signal corresponding to hydroxyl methine carbon (hydroxymethine carbon) and the mesomethylene carbon adjacent with hydroxyl methine carbon (methylene carbon) respectively, thus inference: product Gh-1631 and product Gh-2641-1 are isomers not identical on oh group position, the oh group of product Gh-1631 may be positioned at C-3.

Find from the HMBC spectroscopic data of product Gh-1631: δ 4.75 is associated with δ 168.02 (C-18), δ 105.90 (C-5), δ 75.77 (C-2), δ 75.70 (C-2), δ 25.93 (C-4) and δ 23.54 (C-19); δ 3.07 is associated with δ 90.00 (C-3), δ 105.90 (C-5), δ 75.77 (C-2), δ 75.70 (C-2), 157.32 (C-6), 168.02 (C-18), 103.96 (C-17), 101.72 (C-7) and δ 158.45 (C-16).This proves that oh group is on C-3 position.In addition, because piperazine ring (pyran ring) of muttering has lacked conjugated double bond (conjugated doublebond), it configuration (conformation) is soft form (flexible form), this makes two of C-4 methylene group proton equivalences (equivalence) and has identical chemical shift (chemical shift) [δ 3.07 (2H, d, J=8.1Hz, H-4)], and δ 3.07 and δ 4.75 (1H, t, J=8.1Hz, H-3) coupling.The oh group of this C-3 is axial (axial) (β-), R configuration.

Find from the NOESY spectroscopic data of product Gh-1631: δ 7.42 (H-10) is associated with δ 3.48 (H-11); δ 3.48 (H-11) is associated with δ 2.30 (H1-21); δ 2.53 (H-22) is associated with δ 1.42 (H2-21); δ 2.30 (H1-21) is associated with δ 1.23 (H-24); δ 5.32 (H-27) is associated with δ 1.597 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (carboxylgroup) (C-30) are for trans, and double bond Δ 27,28 is Z configuration.In addition, δ 4.75 (H-3) is associated with δ 3.07 (H-4), δ 1.42 (H-19), δ 1.47 (H2-20) and δ 2.02 (H2-36), this expression C-3 proton is equator is axially (α-) to (α-) and the methyl group be connected on C-2 position, so C-2 is R configuration.

Comprehensive above data, product Gh-1631 is identified is a compounds with the following chemical structure formula:

Product Gh-1631 is named as formoxanthone C (formoxanthone C), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3a S, 5S, 10R, 11R, 14aS)-3a, 4, 5, 7, 10, 11-six hydrogen-8, 10-dihydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 9H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 10R, 11R, 14aS)-3a, 4, 5, 7, 10, 11-hexahydro-8, 10-dihydroxy-3, 3, 11-trimethyl-13-(3-methyl-2-butenyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 9H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

24. product Gh-1050:

Product Gh-1050 is as follows by the character recorded:

Yellow powder, fusing point: 55-57 DEG C.

EIMS m/z (relative intensity): 644 [M]+(82), 616 (98), 601 (9), 598 (8), 571 (16), 533 (17), 517 (12), 490 (100), 489 (96), 475 (19), 447 (30), 433 (13), 405 (18), 371 (33), 363 (17), 309 (10), 295 (21), 253 (25), 230 (19), 213 (15), 189 (9), 173 (10), 147 (11), 105 (17), 99 (25), 69 (42).HREIMS [M]+m/z:644.2983; Calculated value about C38H44O9: 644.2985.

1H-NMR(600MHz，CDCl3)：δ12.69(1H，s，OH-6)，7.47(1H，d，J＝6.8Hz，H-10)，5.57(1H，t，J＝7.5Hz，H-27)，5.19(1H，t，J＝6.8Hz，H-32)，4.63(1H，s，H1-40)，4.29(1H，s，H2-40)，3.86(1H，d，J＝3.4Hz，H-3)，3.65(1H，t，J＝2.6Hz，H-4)，3.49(1H，t，J＝5.7Hz，H-11)，3.31(1H，m，H1-31)，3.28(1H，m，H1-26)，3.23(1H，dd，J＝14.6，6.2Hz，H2-31)，2.82(1H，dd，J＝14.9，5.6Hz，H2-26)，2.55(1H，d，J＝9.4Hz，H-22)，2.31(1H，m，H1-21)，2.29(1H，m，H-37)，2.03(1H，br d，J＝13.3Hz，H1-20)，1.92(3H，s，H-39)，1.76(3H，s，H-34)，1.70(3H，s，H-25)， 1.69(3H，s，H-35)，1.62(3H，s，H-29)，1.60(1H，m，H2-20)，1.49(3H，s，H-19)，1.39(1H，m，H2-21)，1.31(2H，m，H-36)，1.29(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ204.09(C-12)，178.99(C-8)，168.63(C-30)，163.38(C-18)，162.82(C-6)，155.99(C-16)，146.45(C-38)，136.18(C-27)，134.28(C-10，C-9)，131.50(C-33)，128.27(C-28)，122.31(C-32)，109.58(C-40)，106.42(C-17)，100.23(C-5)，100.12(C-7)，90.17(C-14)，84.66(C-23)，83.64(C-13)，79.48(C-2)，71.37(C-3)，48.90(C-22)，48.27(C-37)，47.01(C-11)，38.29(C-20)，36.69(C-4)，29.93(C-25)，29.47(C-26)，28.85(C-24)，25.73(C-35)，25.14(C-21)，23.66(C-19)，22.95(C-39)，21.98(C-36)，21.92(C-31)，20.93(C-29)，18.15(C-34)。

The EIMS data presentation of product Gh-1050: have a part peak [M]+position in m/z 644 and a benchmark peak position at m/z 490 (100), and HREIMS data presentation [M]+m/z 644.2983, this and product Gh-2603-2 are (namely, gambogellic acid) cataclasm graphic (fragmentationpatterns) similar, but more than gambogellic acid 16 mass units (mass units).In addition, 1H-NMR and the 13C-NMR spectrum of product Gh-1050 is similar to product Gh-2603-2 institute tool person haply.

1H-NMR, 13C-NMR and 1H-1H COSY spectroscopic data display of product Gh-1050: have in the structure of product Gh-1050 the lsopropenyl group of monoterpene outer-methene proton [δ 4.63 (H1-40), δ 4.29 (H2-40); δ 109.58 (C-40)], 2 adjacent methylene group [δ 2.03 (1H, br d, J=13.3Hz, H1-20), 1.60 (1H, m, H2-20); δ 38.29 (C-20) and δ 1.31 (2H, m, H-36); δ 21.98 (C-36)], 1 primary hydroxyl groups [δ 3.86 (1H, d, J=3.4Hz, H-3); δ 71.37 (C-3)] and 1 methin groups [δ 3.65 (1H, t, J=2.6Hz, H-4); δ 36.69 (C-4)].In addition, hydroxyl methine protons δ 3.86 (1H, d, J=3.4Hz, H-3) is adjacent with methine protons δ 3.65 (1H, t, J=2.6Hz, H-4) and be coupled mutually.

Find from the HMBC spectroscopic data of product Gh-1050: hydroxyl methine protons [δ 3.86 (1H, d, J=3.1Hz, H-3)] except being associated with 2 level Four carbon [δ 79.48 (C-2) and 100.23 (C-5)], be also associated with 2 methin groups [δ 36.69 (C-4) and δ 48.27 (C-37)]; δ 3.65 (H-4), except being associated with δ 21.98 (C-38), δ 48.27 (C-37), δ 79.48 (C-2), δ 100.23 (C-5), δ 162.82 (C-6) and δ 100.12 (C-7), is also associated with δ 71.37 (C-3).In addition, methene proton [δ 2.03 (H1-20) and δ 1.60 (H2-20)] is associated with δ 79.48 (C-2), δ 48.27 (C-37), δ 23.66 (C-19) and δ 71.37 (C-3); δ 1.49 (H-19) is associated with δ 79.48 (C-2), δ 71.37 (C-3), δ 21.98 (C-36) and aromatic nucleus carbon δ 163.38 (C-18).It can thus be appreciated that oh group is positioned on C-3 position, and monoterpene ring links with the C-18 of ehter bond and aromatic nucleus at C-2, the C-5 of C-4 and aromatic nucleus links.

Find from the NOESY spectroscopic data of product Gh-1050: δ 7.47 (H-10) is associated with δ 3.49 (H-11); δ 3.49 (H-11) is associated with δ 2.31 (H1-21); δ 1.39 (H2-21) is associated with δ 2.55 (H-22); δ 2.31 (H1-21) and δ 2.55 (H-22) is all associated with δ 1.29 (H-24); δ 5.57 (H-27) is associated with δ 1.62 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid or gambogellic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are for trans, and double bond Δ 27,28 is Z configuration.In addition, also find from the NOESY spectroscopic data of product Gh-1050: δ 2.29 (H-37), except being associated with δ 3.65 (H-4) and δ 1.31 (H-36), is also associated with δ 3.86 (H-3) and δ 1.60 (H2-20).This proves that monoterpene ring is the chair form configuration of 1,3-bis-axle, H-37 and H-3 is axially (axial), and methyl group (H-19) and lsopropenyl group be all equator to.Hydroxyl methine protons (H-3) is axially, thus C-3 oh group be equator to α-orientation, C-3 is S configuration.In addition, δ 1.62 (H-29) is not associated with δ 4.63 (H1-40) and δ 4.29 (H2-40), and thus C-2 is R configuration.From above data, the three-dimensional arrangement of monoterpene part is 2R, 3S, 4R, 37S configuration.

Comprehensive above data, product Gh-1050 is identified is a compounds with the following chemical structure formula:

Product Gh-1050 is named as 3 Alpha-hydroxy gambogellic acids (3 α-hydroxygambogellic acid), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10S, 13R, 16aS, 17S)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8, 17-dihydroxyl-3, 3, 13-trimethylammonium-15-(3-methyl-2-butene base)-10-(1-methyl ethylene)-7, 18-dioxy-1, 5:9, 13-dimethylene-1H, 3H, 9H-furo [3.4-g] oxygen is [3.2-b]-1-base also]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 9R, 10S, 13R, 16aS, 17S)-3a, 4, 5, 7, 10, 11, 12, 13-octahydro-8, 17-dihydroxy-3, 3, 13-trimethyl-15-(3-methyl-2-butenyl)-10-(1-methylethenyl)-7, 18-dioxo-1, 5:9, 13-dimethano-1H, 3H, 9H-furo [3.4-g] oxocino [3.2-b] xanthen-1-yl]-, (2Z)-].

25. product Gh-3291:

Product Gh-3291 is as follows by the character recorded:

Yellow powder, fusing point: 103-106 DEG C.

EIMS m/z (relative intensity): 644 [M]+(10), 626 (10), 616 (12), 545 (67), 517 (19), 471 (15), 459 (6), 419 (10), 389 (21), 349 (11), 347 (20), 309 (9), 295 (17), 271 (13), 253 (20), 245 (26), 227 (21), 215 (56), 189 (23), 171 (11), 147 (16), 129 (28), 105 (35), 99 (50), 84 (43), 69 (84), 55 (100).HREIMS [M]+m/z:644.2991; Calculated value about C38H44O9: 644.2985.

1H-NMR(600MHz，CDCl3)：δ12.76(1H，s，OH-6)，7.53(1H，d，J＝7.0Hz，H-10)，6.60(1H，d，J＝10.2Hz，H-4)，6.06(1H，t，J＝7.6Hz，H-27)，5.36(1H，d，J＝10.3Hz，H-3)，5.00(1H，br t，J＝6.3Hz，H-32)，4.89(1H，s，H1-40)，4.81(1H，d，J＝1.0Hz，H2-40)，4.00(1H，t，J＝6.2Hz，H-37)，3.46(1H，dd，J＝6.4，5.1Hz，H-11)，3.28(1H，dd，J＝14.9，8.1Hz，H1-31)，3.13(1H，br dd，J＝14.6，4.7Hz，H2-31)，2.93(1H，dd，J＝16.3，7.9Hz，H1-26)，2.87(1H，dd，J＝16.0，7.6Hz，H2-26)，2.51(1H，d，J＝9.3Hz，H-22)，2.30(1H，dd，J＝13.5，4.7Hz，H1-21)，1.80(1H，m，H1-20)，1.73(3H，s，H-29)，1.71(3H，s，H-25)，1.68(3H，s，H-34)，1.67(1H，m，H1-36)，1.66(3H，m，H-39)，1.62(3H，s，H-35)，1.61(1H，m，H2-36)，1.54(1H，m，H2-20)，1.39(3H，s，H-19)，1.38(1H，m，H2-21)，1.27(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ203.19(C-12)，178.90(C-8)，169.81(C-30)，161.29(C-18)，157.61(C-16)，157.42(C-6)，147.09(C-38)，137.26(C-27)，135.40(C-10)，133.31(C-9)，131.65(C-33)，127.90(C-28)，124.41(C-3)，122.27(C-32)，116.10(C-4)，111.34(C-40)，107.64(C-17)，102.63(C-5)，100.50(C-7)，90.91(C-14)，84.03(C-23)，83.73(C-13)，81.07(C-2)，75.80(C-37)，49.00(C-22)，46.83(C-11)，37.71(C-20)，29.82(C-25)，29.29(C-36)，29.05(C-26)，28.83(C-24)，27.73(C-19)，25.63(C-35)，25.15(C-21)，21.58(C-31)，20.76(C-29)，18.11(C-34)，17.45(C-39)。

The EIMS data presentation of product Gh-3291: have a part peak [M]+position at m/z 644, and HREIMS data presentation [M]+m/z 644.2991, this represents that the molecular formula of product Gh-3291 is C38H44O9, more than gambogic acid 16 mass units.

It is different from gambogic acid institute tool person that 1H-, 13C-NMR and HMQC spectrum of product Gh-3291 shows the side chain be connected on C-2 position, and 3-hydroxy-4-methyl-4-pentenyl (3-hydroxy-4-methyl-4-pentenyl) instead of the 4-methyl-3-pentenyl of gambogic acid.In addition, 1H-NMR and the 13C-NMR spectrum of product Gh-3291 also shows: the structure of product Gh-3291 has 1 primary hydroxyl groups [δ 4.00 (1H, t, J=6.2Hz); δ 75.80] and outer-methylene group [δ 4.89 (1H, s) and δ 4.81 (1H, d); δ 111.34].

Find from the HMBC spectroscopic data of product Gh-3291: hydroxyl methine protons δ 4.00, except being associated with δ 111.34 (C-40) and δ 147.09 (C-38), is also associated with δ 17.45 (C-39), δ 29.29 (C-36) and δ 37.71 (C-20); Methyl proton [δ 1.39 (H-19)], except being associated with δ 37.71 (C-20), is also associated with δ 81.07 (C-2) and δ 124.41 (C-3); δ 5.36 (H-3) is associated with δ 81.07 (C-2), δ 27.73 (C-19) and δ 37.71 (C-20).

The 1H-1H COSY spectroscopic data display of product Gh-3291: δ 4.00 (1H, t, J=6.2Hz, H-37) is coupled with methene proton [δ 1.67 (H1-36), δ 1.61 (H2-36)].This represents that oh group is positioned on C-37 position.

Find from NOESY spectroscopic data: δ 7.53 (H-10) is associated with δ 3.46 (H-11); δ 3.46 (H-11) is associated with δ 2.30 (H1-21); δ 1.38 (H2-21) is associated with δ 2.51 (H-22) and δ 1.27 (H-24); δ 6.06 (H-27) is associated with δ 1.73 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.

In addition, also find from NOESY spectroscopic data: hydroxyl methine protons [δ 4.00 (H-37)] is associated with δ 4.89 (H1-40) and δ 1.66 (H-39); Be associated with the proton [δ 1.80 (H1-20), δ 1.54 (H2-20) and δ 1.67 (H1-36), δ 1.61 (H2-36)] of 2 methylene groups again; δ 5.36 (H-3) is associated with δ 1.39 (H-19), δ 1.80 (H1-20), δ 1.54 (H2-20), δ 1.67 (H1-36) and δ 1.61 (H2-36).In addition, because δ 1.39 (H-19) is associated with δ 1.73 (H-29), so C-2 is R configuration.

Comprehensive above data, product Gh-3291 is identified is a compounds with the following chemical structure formula:

Product Gh-3291 is named as formoxanthone D (formoxanthone D), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(3-hydroxy-4-methyl-4-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(3-methyl-2-butenyl)-11-(3-hydroxy-4-methyl-4-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

26. product Gh-3352:

Product Gh-3352 is as follows by the character recorded:

Yellow powder, fusing point: 194-197 DEG C.

FABMS m/z (relative intensity): 645 [M+H]+(11), 634 (10), 591 (11), 575 (13), 574 (22), 573 (45), 559 (12), 545 (16), 544 (11), 487 (15), 417 (16), 405 (11), 391 (13), 375 (10), 371 (10), 349 (11), 338 (12), 321 (12), 307 (27), 299 (17), 295 (17), 289 (23), 259 (19), 255 (21), 219 (24), 215 (20), 213 (27), 189 (18), 176 (21), 165 (24), 154 (100), 136 (92), 121 (30), 107 (56), 91 (55), 89 (50), 77 (59), 69 (69), 57 (69), 55 (66).HRFABMS [M+H]+m/z:645.3066; Calculated value about C38H45O9: 645.3064.

1H-NMR(600MHz，CDCl3)：δ12.92(1H，s，OH-6)，7.52(1H，d，J＝6.9Hz，H-10)，6.66(1H，d，J＝10.1Hz，H-4)，5.58(1H，br t，J＝6.5Hz，H-27)，5.45(1H，d，J＝10.2Hz，H-3)，5.03(1H，t，J＝7.0Hz，H-37)，3.79(1H，dd，J＝9.2，3.1Hz，H-32)，3.50(1H，dd，J＝6.8，4.7Hz，H-11)，3.30(1H，dd，J＝14.8，10.1Hz，H1-26)，2.90(1H，ddd，J＝15.3，5.6，1.3Hz，H2-26)，2.85(1H，dd，J＝13.9，9.7Hz，H1-31)，2.71(1H，dd，J＝13.8，3.4Hz，H2-31)，2.49(1H，d，J＝9.3Hz，H-22)，2.33(1H，dd，J＝13.5，4.7Hz，H1-21)，2.05(2H，m，H-36)，1.74(3H，s，H-25)，1.73(1H，m，H1-20)，1.69(3H，s，H-29)，1.66(1H，m，H2-20)，1.63(3H，s，H-39)，1.53(3H，s，H-40)，1.45(3H，s，H-19)，1.33(1H，m，H2-21)，1.29(3H，s，H-34)，1.27(3H，s，H-35)，1.25(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ202.89(C-12)，179.05(C-8)，168.89(C-30)，161.34(C-18)，158.49(C-6)，158.28(C-16)，136.09(C-27)，135.01(C-10)，133.19(C-9)，132.17(C-38)，129.31(C-28)，124.75(C-3)，123.46(C-37)，115.93(C-4)，104.83(C-17)，102.92(C-5)，100.67(C-7)，90.77(C-14)，84.28(C-13)，83.86(C-23)，81.81(C-2)，77.20(C-32)，73.24(C-33)，49.12(C-22)，47.04(C-11)，41.72(C-20)，30.56(C-25)，29.65(C-26)，28.89(C-24)，27.18(C-19)，25.89(C-31)，25.69(C-39)，25.63(C-35)，25.36(C-21)，23.75(C-34)，22.66(C-36)，20.73(C-29)，17.59(C-40)。

The data presentation of the HRFABMS of product Gh-3352: have a quasi-molecular ion peak (pseudomolecular ion peak) [M+H]+position at m/z 645.3066, this represents that the molecular formula of product Gh-3352 is C38H44O9, more than gambogic acid institute tool person 16 mass units.

It is different from gambogic acid institute tool person that 1H-, 13C-NMR and HMQC spectrum of product Gh-3352 shows the side chain be connected on C-17 position, 2,3-epoxy group(ing)-3-methyl butyl (2,3-epoxy-3-methylbutyl) instead of the 3-methyl-2-butene base (3-methyl-2-butenyl) of gambogic acid.This represents compared to gambogic acid, and the unsaturated number (unsaturation number) of product Gh-3352 does not change.

1H-1H COSY, HMQC and HMBC spectroscopic data display of product Gh-3352: 1 oxygen methine protons (oxymethine proton) [δ 3.79 (1H, dd, J=9.2,3.1Hz)] with 1 methene proton [δ 2.85 (1H, dd, J=13.9,9.7Hz, H1-31)] coupling; δ 2.85 is coupled with another methene proton [δ 2.71 (1H, dd, J=13.8,3.4Hz, H2-31)] again.In addition, product Gh-3352 contains 2 three grades of methyl groups [δ 1.29 (3H, s, H-34) and δ 1.27 (3H, s, H-35)] be affixed to containing oxygen level Four carbon.From signal [δ 73.24 (C-33)], oxygen methyne (oxymethine) carbon signal [δ 77.20 (CH)] containing oxygen level Four carbon, and C-31 mesomethylene carbon signal [δ 25.89 (CH2)] is known, cycloalkyl groups is between C-32 and C-33 position.

Find from the HMBC spectroscopic data of product Gh-3352: δ 2.85 (H1-31) associates phase with δ 77.20 (C-32) and δ 104.83 (C-17).This proves that the side chain be connected on C-17 position is 2,3-epoxy group(ing)-3-methyl butyl (2,3-epoxy-3-methylbutyl).

Find from the NOESY spectroscopic data of product Gh-3352: δ 7.52 (H-10) is associated with δ 3.50 (H-11); δ 3.50 (H-11) is associated with δ 2.33 (H1-21) and δ 1.33 (H2-21); δ 1.33 (H2-21) is associated with δ 2.49 (H-22); δ 2.49 (H-22) is associated with δ 1.74 (H-25); δ 5.58 (H-27) is associated with δ 1.69 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 1.69 (H-29) is associated with δ 1.45 (H-19), so C-2 is R configuration.

Comprehensive above data, product Gh-3352 is identified is a compounds with the following chemical structure formula:

Product Gh-3352 is named as formoxanthone E (formoxanthone E), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(2, 3-epoxy group(ing)-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(2, 3-epoxy-3-methylbutyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

27. product Gh-3351:

Product Gh-3351 is as follows by the character recorded:

Yellow powder, fusing point: 168-171 DEG C.

EIMS m/z (relative intensity): 621 (6), 603 (17), 589 (100), 577 (23), 561 (19), 503 (47), 467 (6), 423 (8), 381 (5), 339 (4), 315 (15), 231 (7), 213 (9), 135 (4), 69 (15).HRFABMS [M+H]+m/z:645.3070; Calculated value about C38H45O9: 645.3064.

1H-NMR(600MHz，CDCl3)：δ12.91(1H，s，OH-6)，7.51(1H，d，J＝6.9Hz，H-10)，6.49(1H，d，J＝10.1Hz，H-4)，5.56(1H，br t，J＝7.3Hz，H-27)，5.43(1H，d，J＝10.2Hz，H-3)，5.03(1H，t，J＝7.0Hz，H-37)，3.76(1H，br d，J＝7.0Hz，H-32)，3.50(1H，dd，J＝6.6，4.9Hz，H-11)，3.29(1H，dd，J＝15.2，10.1Hz，H1-26)，2.89(1H，ddd，J＝15.3，5.5，1.5Hz，H2-26)，2.82(1H，dd，J＝13.9，9.9Hz，H1-31)，2.69(1H，brd，J＝12.7Hz，H2-31)，2.48(1H，d，J＝9.3Hz，H-22)，2.32(1H，dd，J＝13.5，4.0Hz，H1-21)，2.03(2H，m，H-36)，1.73(3H，s，H-25)，1.68(3H，s，H-29)，1.67(2H，m，H-20)，1.62(3H，s，H-39)，1.53(3H，s，H-40)，1.43(3H，s，H-19)，1.30(1H，m，H2-21)，1.28(3H，s，H-34)，1.26(3H，s，H-35)，1.24(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ202.99(C-12)，179.03(C-8)，169.60(C-30)，161.37(C-18)，158.46(C-6)，158.25(C-16)，135.76(C-27)，134.92(C-10)，133.21(C-9)，132.14(C-38)，129.58(C-28)，124.72(C-3)，123.47(C-37)，115.90(C-4)，104.81(C-17)，102.88(C-5)，100.63(C-7)，90.74(C-14)，84.26(C-13)，83.79(C-23)，81.75(C-2)，77.15(C-32)，73.25(C-33)，49.11(C-22)，47.04(C-11)，41.71(C-20)，30.56(C-25)，29.69(C-26)，28.88(C-24)，27.13(C-19)，25.92(C-31)，25.89(C-39)，25.63(C-35)，25.36(C-21)，23.44(C-34)，22.65(C-36)，20.74(C-29)，17.59(C-40)。

The EIMS data presentation of product Gh-3351: have a benchmark peak position at m/z 589 (100); And HRFABMS data presentation: have a quasi-molecular ion peak [M+H]+position at m/z 645.3070, this represents that the molecular formula of product Gh-3351 is C38H44O9, more than gambogic acid institute tool person 16 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-3351: the side chain be connected on C-17 position is different from gambogic acid institute tool person, and 2,3-epoxy group(ing)-3-methyl butyl instead of the 3-methyl-2-butene base of gambogic acid.This represents compared to gambogic acid, and the unsaturated number of product Gh-3351 does not change.

1H-1H COSY, HMQC and HMBC spectroscopic data display of product Gh-3351: 1 oxygen methine protons δ 3.76 (1H, br d, J=7.0Hz) and 1 methene proton δ 2.82 (1H, dd, J=13.9,9.9Hz, H1-31) coupling; [δ's δ 2.82 2.69 (1H, br d, J=12.7Hz, H2-31) is coupled with another methene proton again.In addition, product Gh-3351 has 2 containing oxygen three grades of methyl protons (oxygen-bearing tertiary methyl protons) [δ 1.28 (3H, s, H-34) and δ 1.26 (3H, s, H-35)].According to the signal containing oxygen level Four carbon [δ 73.25 (C-33)], oxygen methin groups (oxymethine group) [δ 77.15 (C-32)] and methylene group [δ 25.92 (C-31)], cycloalkyl groups is between C-32 and C-33 position.

Find from the HMBC spectroscopic data of product Gh-3351: δ 2.82 (H1-31) is associated with δ 77.15 (C-32), δ 102.88 (C-5), δ 104.81 (C-17), 158.25 (C-16) and δ 161.37 (C-18).This proves that the side chain be connected on C-17 position is 2,3-epoxy group(ing)-3-methyl butyl.

Find from the NOESY spectroscopic data of product Gh-3351: δ 7.51 (H-10) is associated with δ 3.50 (H-11); δ 3.50 (H-11) is associated with δ 2.32 (H1-21) and δ 1.30 (H2-21); δ 1.30 (H2-21) is associated with δ 2.48 (H-22); δ 2.48 (H-22) is associated with δ 1.73 (H-25); δ 5.56 (H-27) is associated with δ 1.68 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are for trans, and double bond Δ 27,28 is Z configuration.In addition, because δ 1.68 (H-29) is associated with δ 2.03 (H-36), so C-2 is S configuration.

Comprehensive above data, product Gh-3351 is identified is a compounds with the following chemical structure formula:

Product Gh-3351 is named as epiformoxanthone E (epiformoxanthone E), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(2, 3-epoxy group(ing)-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-, base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(2, 3-epoxy-3-methylbutyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

28. product Gh-1052:

Product Gh-1052 is as follows by the character recorded:

Yellow powder, fusing point: 83-85 DEG C.

EIMS m/z (relative intensity): 662 [M]+(6), 634 (8), 579 (100), 551 (16), 545 (9), 507 (14), 489 (6), 417 (12), 389 (4), 349 (4), 295 (5), 245 (8), 214.9 (14), 189 (8), 147 (4), 99 (7), 69 (29).HREIMS [M]+m/z:662.3096; Calculated value about C38H46O10: 662.3091.

1H-NMR(600MHz，CDCl3)：δ12.92(1H，s，OH-6)，7.52(1H，d，J＝7.0Hz，H-10)，6.65(1H，d，J＝10.2Hz，H-4)，5.63(1H，t，J＝7.4Hz，H-27)，5.46(1H，d，J＝10.3Hz，H-3)，5.07(1H，t，J＝7.1Hz，H-37)，3.74(1H，dd，J＝10.0，3.4Hz，H-32)，3.49(1H，dd，J＝6.7，4.7Hz，H-11)，3.25(1H，dd，J＝15.2，9.9Hz，H1-26)，2.89(1H，ddd，J＝15.3，5.9，1.7Hz，H2-26)，2.84(1H，dd，J＝13.9，10.1Hz，H1-31)，2.72(1H，dd，J＝13.9，3.4Hz，H2-31)，2.49(1H，d，J＝9.3Hz，H-22)，2.32(1H，dd，J＝13.5，4.7Hz，H1-21)，2.07(2H，m，H-36)，1.79(1H，m，H1-20)，1.73(3H，s，H-25)，1.69(3H，s，H-29)，1.67(1H，m，H2-20)，1.65(3H，s，H-39)，1.57(3H，s，H-40)，1.37(3H，s，H-19)，1.33(1H，dd，J＝13.5，9.6Hz，H2-21)，1.27(3H，s，H-35)，1.26(3H，s，H-24)，1.24(3H，s，H-34)。

13C-NMR(150MHz，CDCl3)：δ202.93(C-12)，179.10(C-8)，169.18(C-30)，161.47(C-18)，158.49(C-6)，158.22(C-16)，136.45(C-27)，135.16(C-10)，133.12(C-9)，132.66(C-38)，129.08(C-28)，124.56(C-3)，123.36(C-37)，116.12(C-4)，104.92(C-17)，102.95(C-5)，100.63(C-7)，90.82(C-14)，84.18(C-13)，83.82(C-23)，81.76(C-2)，77.11(C-32)，73.09(C-33)，49.08(C-22)，47.03(C-11)，41.74(C-20)，30.62(C-25)，29.54(C-26)，28.83(C-24)，27.21(C-19)，25.87(C-35)，25.68(C-31)，25.65(C-39)，25.33(C-21)，23.65(C-34)，22.91(C-36)，20.74(C-29)，17.71(C-40)。

The EIMS data presentation of product Gh-1052: have a part peak [M]+position in m/z 662 and a benchmark peak position at m/z 579 (100), and HREIMS data presentation [M]+m/z 662.3096, this represents that the molecular formula of product Gh-1052 is C38H46O10, more than gambogic acid institute tool person 34 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-1052: the side chain be connected on C-17 position is different from gambogic acid institute tool person, and 2,3-dihydroxyl-3-methyl butyl instead of the 3-methyl-2-butene base of gambogic acid.

1H-NMR and the 13C-NMR spectrum display of product Gh-1052: have 1 primary hydroxyl groups [δ 3.74 (1H, dd, J=10.0,3.4Hz); δ 77.11] and 1 containing oxygen level Four carbon (δ 73.09).In addition, the 1H-1H COSY spectroscopic data display of product Gh-1052: hydroxyl methine protons (hydroxymethine proton) δ 3.74 (1H, dd, J=10.0,3.4Hz) with 2 methene protons (methylene protons) [δ 2.84 (1H, dd, J=13.9,10.1Hz, and δ 2.72 (1H, dd, J=13.9 H1-31), 3.4Hz, H2-31)] coupling.

Find from the HMQC spectroscopic data of product Gh-1052: C-31 (δ 25.68) is methylene group; Primary hydroxyl groups is positioned at C-32 (δ 77.11); Three grades of oh groups are positioned at C-33 (δ 73.09); And C-35 (δ 25.87) is three grades of methyl groups with C-34 (δ 23.65).

Find from the HMBC spectroscopic data of product Gh-1052: δ 3.74 (H-32) is associated with δ 73.09 (C-33) and δ 23.65 (C-34); δ 2.84 (H1-31) and δ 2.72 (H2-31) is all associated with δ 104.92 (C-17), δ 161.47 (C-18), δ 158.22 (C-16) and δ 77.11 (C-32); H1-31 is also associated with δ 73.09 (C-33); δ 1.24 (H-34) and δ 1.27 (H-35) is all associated with δ 77.11 (C-32) and δ 73.09 (C-33); H-34 is also associated with δ 25.87 (C-35); H-35 and δ 23.65 (C-34) is associated.This proves that the side chain be connected on C-17 position is 2,3-dihydroxyl-3-methyl butyl.

Find from the NOESY spectroscopic data of product Gh-1052: δ 7.52 (H-10) is associated with δ 3.49 (H-11); δ 3.49 (H-11) is associated with δ 2.32 (H1-21) and δ 1.33 (H2-21); δ 1.33 (H2-21) is associated with δ 2.49 (H-22); δ 2.49 (H-22) is associated with δ 1.73 (H-25); δ 5.63 (H-27) is associated with δ 1.69 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 1.37 (H-19) is associated with δ 1.69 (H-29), so C-2 is R configuration.

Comprehensive above data, product Gh-1052 is identified is a compounds with the following chemical structure formula:

Product Gh-1052 is named as formoxanthone F (formoxanthone F), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(2, 3-dihydroxyl-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tet rahydro-8-hydroxy-3, 3, 11-trimethyl-13-(2, 3-dihydroxy-3-methylbutyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

29. product Gh-1036:

Product Gh-1036 is as follows by the character recorded:

Yellow powder, fusing point: 76-78 DEG C.

EIMS m/z (relative intensity): 662 [M]+(13), 634 (7), 579 (100), 551 (13), 545 (10), 507 (12), 489 (5), 417 (9), 375 (3), 349 (3), 295 (4), 245 (8), 215 (8), 213 (7), 147 (4), 105 (5), 69 (19).HREIMS [M]+m/z662.3098; Calculated value about C38H46O10: 662.3091.

1H-NMR(600MHz，CDCl3)：δ12.91(1H，s，OH-6)，7.52(1H，d，J＝6.9Hz，H-10)，6.66(1H，d，J＝10.0Hz，H-4)，5.53(1H，br t，J＝5.2Hz，H-27)，5.47(1H，d，J＝10.1Hz，H-3)，5.08(1H，t，J＝6.9Hz，H-37)，3.74(1H，dd，J＝11.2，2.6Hz，H-32)，3.49(1H，t，J＝5.6Hz，H-11)，3.27(1H，dd，J＝15.0，10.4Hz，H1-26)，2.87(1H，dd，J＝13.6，11.7Hz，H1-31)，2.82(1H，br dd，J＝15.3，3.8Hz，H2-26)，2.72(1H，dd，J＝13.7，2.8Hz，H2-31)，2.48(1H，d，J＝9.2Hz，H-22)，2.32(1H，dd，J＝13.5，4.5Hz，H1-21)，2.10(2H，m，H-36)，1.78(1H，m，H1-20)，1.702(3H，s，H-25)，1.697(1H，m，H2-20)，1.653(3H，s，H-29)，1.648(3H，s，H-39)，1.58(3H，s，H-40)，1.35(3H，s，H-19)，1.33(1H，m，H2-21)，1.28(3H，s，H-35)，1.25(3H，s，H-24)，1.24(3H，s，H-34)。

13C-NMR(150MHz，CDCl3)：δ202.96(C-12)，179.04(C-8)，168.66(C-30)，161.45(C-18)，158.35(C-6)，158.28(C-16)，135.28(C-27)，134.98(C-10)，133.19(C-9)，132.39(C-38)，129.46(C-28)，124.77(C-3)，123.52(C-37)，116.07(C-4)，104.54(C-17)，103.19(C-5)，100.64(C-7)，90.87(C-14)，84.28(C-13)，83.60(C-23)，81.71(C-2)，76.57(C-32)，73.06(C-33)，49.07(C-22)，46.99(C-11)，41.68(C-20)，30.76(C-25)，29.96(C-26)，28.86(C-24)，26.79(C-19)，26.29(C-35)，25.63(C-39)，25.42(C-21)，24.62(C-31)，23.56(C-34)，22.82(C-36)，20.52(C-29)，17.62(C-40)。

The EIMS data presentation of product Gh-1036: have a part peak [M]+position in m/z 662 and a benchmark peak position at m/z 579 (100), and HREIMS data presentation [M]+m/z 662.3098, this represents that the molecular formula of product Gh-1036 is C38H46O10, more than gambogic acid institute tool person 34 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-1036: the side chain be connected on C-17 position is different from gambogic acid institute tool person, and 2,3-dihydroxyl-3-methyl butyl instead of the 3-methyl-2-butene base of gambogic acid.

1H-and the 13C-NMR spectrum display of product Gh-1036: have 1 primary hydroxyl groups [δ 3.74 (1H, dd, J=11.2,2.6Hz); δ 76.57] and 1 containing oxygen level Four carbon (δ 73.06).

In addition, the 1H-1H COSY spectroscopic data display of product Gh-1036: hydroxyl methine protons δ 3.74 (1H, dd, J=11.2,2.6Hz) with 2 methene protons [δ 2.87 (1H, dd, J=13.6,11.7Hz, and δ 2.72 (1H, dd, J=13.7 H1-31), 2.8Hz, H2-31)] coupling.

Find from the HMQC spectroscopic data of product Gh-1036: C-31 (δ 24.62) is methylene group ` primary hydroxyl groups is be positioned at C-32 (δ 76.57); Three grades of oh groups are positioned at C-33 (δ 73.06); And C-34 (δ 23.56) is three grades of methyl groups with C-35 (δ 26.29).

Find from the HMBC spectroscopic data of product Gh-1036: δ 3.74 (H-32), except being associated with δ 24.62 (C-31), δ 73.06 (C-33) and δ 23.56 (C-34), is also associated with δ 104.54 (C-17); δ 2.87 (H1-31) and δ 2.72 (H2-31) is all associated with δ 76.57 (C-32), δ 73.06 (C-33), δ 104.54 (C-17), δ 161.45 (C-18) and δ 158.28 (C-16).This proves that the side chain be connected on C-17 position is 2,3-dihydroxyl-3-methyl butyl.

Find from the NOESY spectroscopic data of product Gh-1036: δ 7.52 (H-10) is associated with δ 3.49 (H-11); δ 3.49 (H-11) is associated with δ 2.32 (H1-21) and δ 1.33 (H2-21); δ 1.33 (H2-21) is associated with δ 2.48 (H-22); δ 2.48 (H-22) is associated with δ 1.702 (H-25); δ 5.53 (H-27) is associated with δ 1.653 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 1.653 (H-29) is associated with δ 1.78 (H1-20) and δ 2.10 (H-36), so C-2 is S configuration.

Comprehensive above data, product Gh-1036 is identified is a compounds with the following chemical structure formula:

Product Gh-1036 is named as epiformoxanthone F (epiformoxanthone F), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(2, 3-dihydroxyl-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(2, 3-dihydroxy-3-methylbutyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

30. product Gh-3353:

Product Gh-3353 is as follows by the character recorded:

Yellow powder, fusing point: 158-162 DEG C.

EIMS m/z (relative intensity): 590 (32), 589 (100), 577 (24), 561 (19), 503 (44), 467 (6), 423 (8), 381 (5), 339 (4), 315 (8), 3--(3), 285 (2), 247 (4), 231 (6), 205 (2), 135 (4), 81 (3), 69 (14).HRFABMS [M+H]+m/z:661.3019; Calculated value about C38H45O10: 661.3013.

1H-NMR(600MHz，CDCl3)：δ13.13(1H，s，OH-6)，7.56(1H，d，J＝7.1Hz，H-10)，6.62(1H，d，J＝10.1Hz，H-4)，5.44(1H，d，J＝10.3Hz，H-3)，5.15(1H，d，J＝8.9Hz，H-31)，5.04(1H，br t，J＝3.5Hz，H-37)，5.01(1H，m，H-27)，4.70(1H，d，J＝8.9Hz，H-32)，3.55(1H，dd，J＝6.6，4.9Hz，H-11)，3.42(1H，t，J＝13.4Hz，H1-26)，2.73(1H，br d，J＝13.9Hz，H2-26)，2.52(1H，d，J＝9.3Hz，H-22)，2.36(1H，dd，J＝13.5，7.7Hz，H1-21)，2.05(2H，m，H-36)，1.82(3H，s，H-25)，1.76(1H，m，H1-20)，1.64(3H，s，H-35)，1.63(1H，m，H2-20)，1.62(3H，s，H-39)，1.54(3H，s，H-40)，1.49(3H，s，H-29)，1.44(6H，s，H-19，H-34)，1.27(1H，m，H2-21)，1.26(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ202.26(C-12)，179.10(C-8)，166.33(C-30)，161.08(C-18)，159.90(C-16)，158.73(C-6)，134.79(C-10)，132.77(C-9)，132.73(C-28)，132.09(C-38)，128.48(C-27)，124.60(C-3)，123.55(C-37)，115.48(C-4)，105.98(C-17)，101.92(C-5)，99.90(C-7)，91.89(C-14)，84.63(C-13)，83.75(C-33)，83.70(C-23)，82.16(C-2)，76.83(C-32)，67.26(C-31)，49.47(C-22)，46.71(C-11)，41.97(C-20)，31.14(C-26)，30.13(C-25)，29.42(C-24)，27.60(C-19)，26.05(C-21)，25.63(C-39)，23.89(C-35)，22.43(C-36)，19.98(C-29)，18.80(C-34)，17.66(C-40)。

The EIMS data presentation of product Gh-3353: have a benchmark peak position at m/z 589 (100), and HRFABMS data presentation: have a quasi-molecular ion peak [M+H]+position at m/z 661.3019, this represents that the molecular formula of product Gh-3353 is C38H44O10, more than gambogic acid institute tool person 32 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-3353: the side chain be connected on C-17 position is different from gambogic acid institute tool person, and 1-hydroxyl-2,3 epoxy group(ing)-3-methyl butyl instead of the 3-methyl-2-butene base of gambogic acid.

1H-, 13C-NMR and 1H-1H COSY spectroscopic data display of product Gh-3353: hydroxyl methine protons [δ 5.15 (1H, d, J=8.9Hz)] with 1 the oxygen methine protons [δ 4.70 (1H be positioned in cycloalkyl groups, d, J=8.9Hz)] intercouple.Because the coupling constant (J) of the oxygen methine protons (vicinal oxymethine protons) near 2 on oxirane ring (epoxy ring) is less than 5Hz usually, so the coupling of J=8.9Hz should not be the coupling of the oxygen methine protons near on oxirane ring (epoxy ring) 2.

Find from the HMBC spectroscopic data of product Gh-3353: δ 5.15 is associated with δ 161.08 (C-18), δ 159.90 (C-16), δ 105.98 (C-17) and δ 76.83 (C-32); δ 4.70 is associated with δ 83.75 (C-33), δ 18.80 (C-34), δ 23.89 (C-35) and δ 67.26 (C-31).This proves that oh group is positioned at C-31, and cycloalkyl groups is between C-32 and C-33, and the side chain be connected on C-17 position is 1-hydroxyl-2,3 epoxy group(ing)-3-methyl butyl.

Find from the NOESY spectroscopic data of product Gh-3353: δ 7.56 (H-10) is associated with δ 3.55 (H-11); δ 3.55 (H-11) is associated with δ 2.36 (H1-21) and δ 1.27 (H2-21); δ 2.36 (H1-21) and δ 1.27 (H2-21) is all associated with δ 2.52 (H-22); δ 2.52 (H-22) is associated with δ 1.82 (H-25) and δ 1.26 (H-24); δ 5.01 (H-27) is associated with δ 1.49 (H-29); δ 1.49 (H-29) is not associated with δ 2.05 (H-36) or δ 1.76 (H1-20).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, δ 5.04 (H-37) is associated with δ 1.62 (H-39) and δ 1.54 (H-40); δ 2.05 (H-36) is associated with δ 1.76 (H1-20).Because δ 1.49 (H-29) is not associated with δ 2.05 (H-36) or δ 1.76 (H1-20), so C-2 is R configuration.

Comprehensive above data, product Gh-3353 is identified is a compounds with the following chemical structure formula:

Product Gh-3353 is named as formoxanthone G (formoxanthone G), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14a S)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(1-hydroxyl-2, 3-epoxy group(ing)-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(1-hydroxy-2, 3-epoxy-3-methylbutyl)-11-(4-methy l-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

31. product Gh-3311:

Product Gh-3311 is as follows by the character recorded:

Yellow powder, fusing point: 144-148 DEG C.

EIMS m/z (relative intensity): 590 (7), 589 (18), 577 (5), 561 (9), 503 (11), 467 (2), 423 (2), 347 (5), 315 (3), 285 (3), 247 (6), 233 (13), 231 (22), 230 (16), 215 (8), 202 (4), 131 (6), 117 (100), 115 (19), 91 (12), 69 (14).HRFABMS [M+H]+m/z 661.3010; Calculated value about C38H45O10: 661.3013.

1H-NMR(600MHz，CDCl3)：δ13.12(1H，s，OH-6)，7.56(1H，d，J＝7.1Hz，H-10)，6.65(1H，d，J＝10.0Hz，H-4)，5.52(1H，d，J＝10.2Hz，H-3)，5.17(1H，d，J＝9.0Hz，H-31)，5.05(1H，m，H-37)，5.03(1H，m，H-27)，4.71(1H，d，J＝9.0Hz，H-32)，3.56(1H，dd，J＝6.8，4.8Hz，H-11)，3.42(1H，t，J＝13.5Hz，H1-26)，2.74(1H，br d，J＝13.7Hz，H2-26)，2.54(1H，d，J＝9.3Hz，H-22)，2.37(1H，dd，J＝13.5，4.8Hz，H1-21)，2.05(2H，m，H-36)，1.86(1H，m，H1-20)，1.83(3H，s，H-25)，1.69(1H，m，H2-20)，1.66(3H，s，H-39)，1.64(3H，s，H-35)，1.56(3H，s，H-40)，1.49(3H，s，H-29)，1.45(3H，s，H-34)，1.35(3H，s，H-19)，1.28(1H，m，H2-21)，1.26(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ202.22(C-12)，179.25(C-8)，166.30(C-30)，160.99(C-18)，159.85(C-16)，158.80(C-6)，134.86(C-10)，132.81(C-9)，132.75(C-28)，132.17(C-38)，128.54(C-27)，125.25(C-3)，123.50(C-37)，116.08(C-4)，106.57(C-17)，102.68(C-5)，100.01(C-7)，91.92(C-14)，84.65(C-13)，83.80(C-33)，83.66(C-23)，81.92(C-2)，76.98(C-32)，67.16(C-31)，49.47(C-22)，46.75(C-11)，41.64(C-20)，31.16(C-26)，30.15(C-25)，29.43(C-24)，26.90(C-19)，26.08(C-21)，25.67(C-39)，23.83(C-35)，23.21(C-36)，19.89(C-29)，18.91(C-34)，17.72(C-40)。

The EIMS data presentation of product Gh-3311: have a benchmark peak position at m/z 117 (100), but do not have molecular ion peak [M]+; And HRFABMS data presentation has a quasi-molecular ion peak [M+H]+position at m/z 661.3010, this represents that the molecular formula of product Gh-3311 is C38H44O10, more than gambogic acid institute tool person 32 mass units, and unsaturated number does not change.

1H-NMR and the 13C-NMR spectrum display of product Gh-3311: have 1 primary hydroxyl groups [δ 5.17 (1H, d, J=9.0Hz); δ 67.16] and 1 cycloalkyl groups [δ 4.71 (1H, d, J=9.0Hz); δ 76.98].This spectroscopic data and Gh-3353 institute tool person very similar.

The 1H-1H COSY spectroscopic data display of product Gh-3311: hydroxyl methine protons δ 5.17 (1H, d, J=9.0Hz) with oxygen methine protons δ 4.71 (1H, the d of cycloalkyl groups, J=9.0Hz) intercouple, its coupling constant (J) is 9.0Hz.Because the coupling constant (J) of the oxygen methine protons (vicinal oxymethine protons) near 2 on oxirane ring (epoxy ring) is less than 5Hz usually, so the coupling of J=9Hz should not be the coupling of the oxygen methine protons near on oxirane ring (epoxy ring) 2.

Find from the HMBC spectroscopic data of product Gh-3311: δ 5.17 is associated with δ 160.99 (C-18), δ 159.85 (C-16), δ 106.57 (C-17) and δ 76.98 (C-32); δ 4.71 is associated with δ 83.80 (C-33), δ 18.91 (C-34) and δ 67.16 (C-31).This proves that oh group is positioned at C-31, and cycloalkyl groups is between C-32 and C-33, and the side chain be connected on C-17 position is 1-hydroxyl-2,3 epoxy group(ing)-3-methyl butyl.

Find from the NOESY spectroscopic data of product Gh-3311: δ 7.56 (H-10) is associated with δ 3.56 (H-11); δ 3.56 (H-11) is associated with δ 2.37 (H1-21) and δ 1.28 (H2-21); δ 2.37 (H1-21) and δ 1.28 (H2-21) is all associated with δ 2.54 (H-22); δ 2.54 (H-22) is associated with δ 1.83 (H-25) and δ 1.26 (H-24); δ 5.03 (H-27) is associated with δ 1.49 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 1.49 (H-29) is associated with δ 1.86 (H1-20) and δ 1.69 (H2-20), so C-2 is S configuration.

Comprehensive above data, product Gh-3311 is identified is a compounds with the following chemical structure formula:

Product Gh-3311 is named as piformoxanthone G (epiformoxanthone G), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(1-hydroxyl-2, 3-epoxy group(ing)-3-methyl butyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethy l-13-(1-hydroxy-2, 3-epoxy-3-methylbutyl)-11-(4-methyl-3-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

32. product Gh-3272:

Product Gh-3272 is as follows by the character recorded:

Yellow powder, fusing point: 190-193 DEG C.

EIMS m/z (relative intensity): 626 [M-H2O]+(2), 598 (2), 545 (100), 517 (3), 499 (2), 389 (3), 347 (3), 271 (2), 245 (4), 215 (11), 189 (4), 147 (1), 105 (2), 69 (2).HRFABMS [M-H2O+H]+m/z:627.2961; Calculated value about C38H43O8: 627.2958.

1H-NMR(600MHz，CDCl3)：δ12.71(1H，s，OH-6)，7.52(1H，d，J＝6.9Hz，H-10)，6.57(1H，d，J＝10.1Hz，H-4)，6.03(1H，d t，J＝7.5，1.3Hz，H-27)，5.59(1H，d，J＝15.7Hz，H-37)，5.53(1H，dt，J＝15.6，6.9Hz，H-36)，5.34(1H，d，J＝10.1Hz，H-3)，5.02(1H，d t，J＝6.2，1.2Hz，H-32)，3.46(1H，dd，J＝6.7，4.6Hz，H-11)，3.27(1H，dd，J＝14.7，8.2Hz，H1-31)，3.11(1H，br dd，J＝14.6，5.2Hz，H2-31)，2.94(2H，br t，J＝5.6Hz，H-26)，2.49(1H，d，J＝9.3Hz，H-22)，2.36(1H，dd，J＝13.9，7.2Hz，H1-20)，2.29(1H，dd，J＝13.6，4.8Hz，H1-21)，2.25(1H，dd，J＝14.0，6.5Hz，H2-20)，1.72(3H，s，H-34)，1.70(3H，d，J＝1.1Hz，H-29)，1.67(3H，s，H-25)，1.63(3H，s，H-35)，1.38(3H，s，H-19)，1.37(1H，m，H2-21)，1.27(3H，s，H-24)，1.16(6H，s，H-39、H-40)。

13C-NMR(150MHz，CDCl3)：δ203.30(C-12)，178.89(C-8)，170.80(C-30)，161.41(C-18)，157.41(C-16)，157.37(C-6)，142.05(C-37)，137.79(C-27)，135.37(C-10)，133.30(C-9)，131.51(C-33)，127.80(C-28)，123.91(C-3)，122.23(C-32)，120.93(C-36)，116.30(C-4)，107.41(C-17)，102.97(C-5)，100.39(C-7)，90.90(C-14)，83.90(C-23)，83.78(C-13)，80.84(C-2)，70.61(C-38)，48.95(C-22)，46.81(C-11)，44.67(C-20)，29.83(C-25)，29.44(C-39)，29.43(C-40)，29.27(C-26)，28.83(C-24)，27.39(C-19)，25.73(C-35)，25.14(C-21)，21.58(C-31)，20.72(C-29)，18.14(C-34)。

The EIMS data presentation of product Gh-3272: have one [M-H2O]+(2) position in m/z 626 and a benchmark peak position at m/z 545 (100), and HRFABMS data presentation has a quasi-molecular ion peak [M-H2O+H]+position at m/z 627.2961, this represents that the molecular formula of product Gh-3272 is C38H44O9, more than gambogic acid institute tool person 16 mass units, and unsaturated number does not change.

1H-NMR and the 13C-NMR spectrum display of product Gh-3272: the side chain be connected on C-2 position is different from gambogic acid institute tool person, and 4-hydroxy-4-methyl-pentenyl instead of the 4-methyl-3-pentenyl of gambogic acid.

1H-NMR and the 13C-NMR spectrum display of product Gh-3272: product Gh-3272 has 1 containing oxygen level Four carbon (δ 70.61) and 2 thiazolinyl proton [δ 5.59 (1H be coupled mutually, d, and δ 5.53 (1H J=15.7Hz), dt, J=15.6,6.9Hz)].Because coupling constant (J) is 15.7Hz, so thiazolinyl is trans double bond (namely E configuration).

The 1H-1H COSY spectroscopic data display of product Gh-3272: δ 5.53 (1H, dt, J=15.6,6.9Hz) except with δ 5.59 (1H, d, J=15.7Hz), outside coupling, be also coupled with 2 methene protons [δ 2.36 (H1-20) and δ 2.25 (H2-20)].This proves that two replacement trans double bond are that position is at C-36/C-37.

Find from the HMBC spectroscopic data of product Gh-3272: methene proton [δ 2.25 (H2-20) and δ 2.36 (H1-20)] is associated with δ 27.39 (C-19), δ 80.84 (C-2), δ 120.93 (C-36) and δ 142.05 (C-37), and the thiazolinyl carbon δ 123.91 (C-3) muttered with piperazine again in ring is associated; δ 5.53 (H-36) is associated with δ 44.67 (C-20), δ 70.61 (C-38) and δ 142.05 (C-37); δ 5.59 (H-37) is associated with δ 44.67 (C-20), δ 70.61 (C-38), δ 120.93 (C-36), δ 29.44 (C-39) and δ 29.43 (C-40).This proves that the side chain be connected on C-2 position is 4-hydroxy-4-methyl-pentenyl.

Find from the NOESY spectroscopic data of product Gh-3272: δ 7.52 (H-10) is associated with δ 3.46 (H-11); δ 3.46 (H-11) is associated with δ 2.29 (H1-21) and δ 1.37 (H2-21); δ 1.37 (H2-21) is associated with δ 2.49 (H-22); δ 2.49 (H-22) is associated with together with-dimethyl group (gem-dimethyl group) [δ 1.67 (H-25) and δ 1.27 (H-24)]; δ 6.03 (H-27) is associated with δ 1.70 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, thiazolinyl proton [δ 5.53 (H-36)], except being associated with δ 5.59 (H-37) and methene proton [δ 2.36 (H1-20), δ 2.25 (H2-20)], also with together with-dimethyl group [δ 1.16 (H-39, H-40)] is associated; Another thiazolinyl proton [δ 5.34 (H-3)] is associated with δ 1.38 (H-19) and δ 2.25 (H2-20).Because δ 1.38 (H-19) is associated with δ 1.70 (H-29), so C-2 is R configuration.

Comprehensive above data, product Gh-3272 is identified is a compounds with the following chemical structure formula:

Product Gh-3272 is named as formoxanthone H (formoxanthone H), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(4-hydroxy-4-methyl-2E-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethy l-13-(3-methyl-2-butenyl)-11-(4-hydroxy-4-methyl-2E-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

33. product Gh-3332:

Product Gh-3332 is as follows by the character recorded:

Yellow powder, fusing point: 102-106 DEG C.

EIMS m/z (relative intensity): 626 [M-H2O]+(5), 575 (2), 545 (100), 499 (2), 471 (1), 389 (4), 347 (4), 271 (3), 245 (5), 215 (15), 189 (5), 147 (2), 105 (3), 69 (2).HRFABMS [M-H2O+H]+m/z:627.2966; Calculated value about C38H43O8: 627.2958.

1H-NMR(600MHz，CDCl3)：δ12.81(1H，s，OH-6)，7.55(1H，d，J＝6.9Hz，H-10)，6.83(1H，dt，J＝7.4，1.3Hz，H-27)，6.64(1H，J＝10.0Hz，H-4)，5.68(1H，d，J＝8.4Hz，H-37)，5.67(1H，m，H-36)，5.44(1H，d，J＝10.0Hz，H-3)，4.97(1H，br t，J＝6.2Hz，H-32)，3.48(1H，dd，J＝6.8，4.5Hz，H-11)，3.28(1H，dd，J＝14.9，8.9Hz，H1-31)，3.14(1H，br dd，J＝13.5，3.4Hz，H2-31)，2.69(1H，ddd，J＝16.4，6.2，1.3Hz，H1-26)，2.52(1H，d，J＝9.3Hz，H-22)，2.42(1H，dd，J＝14.3，4.8Hz，H1-21)，2.32(2H，m，H-20)，2.20(1H，dd，J＝16.3，8.5Hz，H2-26)，1.71(3H，s，H-34)，1.69(3H，s，H-25)，1.61(3H，s，H-35)，1.38(3H，s，H-19)，1.37(3H，s，H-39)，1.36(1H，m，H2-21)，1.33(3H，s，H-40)，1.30(3H，s，H-29)，1.27(3H，s，H-24)。

13C-NMR(150MHz，CDCl3)：δ203.38(C-12)，178.82(C-8)，169.45(C-30)，161.28(C-18)，157.69(C-16)，157.35(C-6)，141.00(C-37)，136.22(C-27)，135.59(C-10)，133.32(C-9)，131.48(C-33)，128.43 (C-28)，125.06(C-3)，122.22(C-32)，121.48(C-36)，116.22(C-4)，108.22(C-17)，102.82(C-5)，100.33(C-7)，91.06(C-14)，83.85(C-23)，83.55(C-13)，80.41(C-2)，71.65(C-38)，49.14(C-22)，46.72(C-11)，43.61(C-20)，29.84(C-25)，29.53(C-39)，29.23(C-40)，28.81(C-24)，28.63(C-26)，26.48(C-19)，25.74(C-35)，24.95(C-21)，21.70(C-31)，18.16(C-34)，11.96(C-29)。

The EIMS data presentation of product Gh-3332: have one [M-H2O]+(5) position in m/z 626 and a benchmark peak position at m/z 545 (100), and HRFABMS data presentation has a quasi-molecular ion peak [M-H2O+H]+position at m/z 627.2966, this represents that the molecular formula of product Gh-3332 is identical with product Gh-3272 institute tool person, be all C38H44O9, and than gambogic acid or many 16 mass units of Isogambogic acid institute tool person, and unsaturated number does not change.

EIMS, 1H-NMR and 13C-NMR spectroscopic data of product Gh-3332 is similar to product Gh-3272 institute tool person haply, and this represents that product Gh-3332 should be the isomer of product Gh-3272.In addition, 1H-NMR spectroscopic data [δ 1.33 (3H, the s of substituted radical (substituent groups) that will be connected on C-20 position, H-40), δ 1.37 (3H, s, H-39), δ 5.68 (1H, d, J=8.4Hz, H-37) and δ .67 (1H, m, H-36) do one with product Gh-3272 institute tool person after find, the Δ 36,37 of product Gh-3332 is Z configuration.Moreover, with product Gh-3272 in comparison, H-27 has 1 downfield shift shown (downfield shift) (δ H 6.83, Δ δ=0.8), and the impact that C-29 is subject to γ-effect (γ-effect) has 1 upfield shift (upfield shift) (δ C 11.96, Δ δ=-8.76), infer that the Δ 27,28 of product Gh-3332 is E configuration thus.

Comprehensive above data, product Gh-3332 is identified is a compounds with the following chemical structure formula:

Product Gh-3332 is named as isoformoxanthone I (isoformoxanthone I), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(4-hydroxy-4-methyl-2Z-pentenyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2E)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(3-methyl-2-butenyl)-11-(4-hydroxy-4-methyl-2Z-pentenyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2E)-].

34. product Gh-3261:

Product Gh-3261 is as follows by the character recorded:

Yellow powder, fusing point: 103-105 DEG C.

EIMS m/z (relative intensity): 662 [M]+(15), 647 (5), 603 (2), 545 (100), 517 (12), 499 (2), 389 (4), 347 (3), 295 (1), 245 (3), 215 (6), 189 (2), 147 (1), 105 (2), 69 (3), 59 (3).HREIMS [M]+m/z:662.3098; Calculated value about C38H46O10: 662.3091.

1H-NMR(600MHz，CDCl3)：δ12.73(1H，s，OH-6)，7.49(1H，d，J＝7.0Hz，H-10)，6.65(1H，dd，J＝10.2，2.5Hz，H-4)，5.54(1H，br t，J＝4.6Hz，H-27)，5.36(1H，d，J＝10.3Hz，H-3)，5.06(1H，br s，H-32)，3.50(1H，t，J＝5.7Hz，H-11)，3.39(1H，m，H1-26)，3.36(1H，d，J＝10.1Hz，H-37)，3.27(1H，dd，J＝14.9，7.4Hz，H1-31)，3.22(1H，br dd，J＝14.3，4.7Hz，H2-31)，2.86(1H，br d，J＝14.1Hz，H2-26)，2.51(1H，d，J＝9.4Hz，H-22)，2.33(1H，dd，J＝13.5，4.7Hz，H1-21)，2.04(1H，m，H1-20)，1.74(1H，m，H1-36)，1.72(3H，s，H-34)，1.68(1H，m，H2-20)，1.65(3H，s，H-25)，1.64(3H，s，H-35)，1.60(3H，s，H-29)，1.459(1H，m，H2-36)，1.456(3H，s，H-19)，1.39(1H，dd，J＝13.5，9.6Hz，H2-21)，1.27(3H，s，H-24)，1.21(3H，s，H-39)，1.11(3H，s，H-40)。

13C-NMR(150MHz，CDCl3)：δ203.34(C-12)，179.20(C-8)，168.78(C-30)，161.13(C-18)，157.75(C-16)，157.43(C-6)，136.34(C-27)，134.74(C-10)，133.91(C-9)，131.41(C-33)，128.56(C-28)，124.18(C-3)，122.33(C-32)，116.18(C-4)，107.80(C-17)，102.49(C-5)，100.63(C-7)，90.57(C-14)，84.03(C-13)，83.64(C-23)，81.24(C-2)，78.23(C-37)，73.70(C-38)，49.01(C-22)，47.02(C-11)，38.73(C-20)，29.92(C-25)，29.87(C-26)，29.05(C-24)，27.88(C-19)，26.70(C-39)，25.70(C-35)，25.28(C-36)，25.25(C-21)，24.28(C-40)，21.71(C-31)，20.85(C-29)，18.22(C-34)。

The EIMS data presentation of product Gh-3261: have a part peak [M]+position in m/z 662 and a benchmark peak position at m/z 545 (100), and HREIMS data presentation [M]+m/z 662.3098, this represents that the molecular formula of product Gh-3261 is C38H46O10, more than gambogic acid 34 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-3261: the side chain be connected on C-2 position is different from gambogic acid institute tool person, 3,4-dihydroxyl-4-methyl amyl (3,4-dihydroxy-4-methylpentyl) instead of the 4-methyl-3-pentenyl of gambogic acid.In addition, 1H-NMR and the 13C-NMR spectrum display of product Gh-3261: have 1 primary hydroxyl groups [δ 3.36 (1H, d, J=10.1Hz); δ 78.23] and 1 containing oxygen level Four carbon (δ 73.70).

The 1H-1H COSY spectroscopic data display of product Gh-3261: hydroxyl methine protons [δ 3.36 (1H, d, J=10.1Hz)] and δ 1.459 (1H, m) are coupled; δ 1.459 (1H, m) and δ 2.04 (1H, m, H1-20) are coupled; 2 on C-20 and C-36 position adjacent methylene group [δ 2.04 (H1-20), δ 1.68 (H2-20); δ 38.73 (C-20) and δ 1.74 (H1-36), δ 1.459 (H2-36); δ 25.28 (C-36)] 4 protons be coupled mutually.This represents that hydroxyl methine protons is H-37.It can thus be appreciated that primary hydroxyl groups is positioned at C-37 (δ 78.23); Three grades of oh groups are positioned at C-38 (δ 73.70); And C-39 (δ 26.70) is three grades of methyl groups with C-40 (δ 24.28).

Find from the HMBC spectroscopic data of product Gh-3261: hydroxyl methine protons δ 3.36 (H-37) is associated with δ 38.73 (C-20); δ 2.04 (H1-20) and δ 1.68 (H2-20) is all associated with δ 25.28 (C-36), δ 81.24 (C-2) and δ 124.18 (C-3); δ 5.36 (H-3) is associated with δ 102.49 (C-5), δ 81.24 (C-2) and δ 27.88 (C-19); δ 1.456 (H-19) is associated with δ 81.24 (C-2), δ 124.18 (C-3), δ 116.18 (C-4) and δ 38.73 (C-20); δ 1.21 (H-39) is associated with δ 78.23 (C-37), δ 73.70 (C-38) and δ 24.28 (C-40); δ 1.11 (H-40) is associated with δ 78.23 (C-37), δ 73.70 (C-38) and δ 26.70 (C-39).This proves that the side chain be connected on C-2 position is 3,4-dihydroxyl-4-methyl amyl.

Find from the NOESY spectroscopic data of product Gh-3261: δ 7.49 (H-10) is associated with δ 3.50 (H-11); δ 3.50 (H-11) is associated with δ 2.33 (H1-21) and δ 1.39 (H2-21); δ 1.39 (H2-21) is associated with δ 2.51 (H-22) and δ 1.27 (H-24); δ 2.51 (H-22) is associated with δ 1.65 (H-25); δ 5.54 (H-27) is associated with δ 1.60 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 3.36 (H-37) is associated with δ 1.21 (H-39) and δ 1.11 (H-40), and δ 1.456 (H-19) is associated with δ 1.60 (H-29), so C-2 is R configuration.

Comprehensive above data, product Gh-3261 is identified is a compounds with the following chemical structure formula:

Product Gh-3261 is named as formoxanthone J (formoxanthone J), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(3, 4-dihydroxyl-4-methyl amyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11R, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(3-methyl-2-butenyl)-11-(3, 4-dihydroxy-4-methylpentyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

35. product Gh-3271:

Product Gh-3271 is as follows by the character recorded:

Yellow powder, fusing point: 99-102 DEG C.

EIMS m/z (relative intensity): 662 [M]+(14), 644 (5), 603 (4), 545 (100), 517 (15), 499 (3), 419 (4), 389 (6), 347 (5), 283 (5), 245 (6), 215 (11), 213 (5), 189 (5), 147 (4), 129 (4), 117 (5), 105 (8), 91 (7), 85.9 (12), 83.9 (19), 69 (11), 59 (8), 57 (11), 55 (11).HREIMS [M]+m/z:662.3097; Calculated value about C38H46O10: 662.3091.

1H-NMR(600MHz，CDCl3)：δ12.75(1H，s，OH-6)，7.50(1H，d，J＝7.0Hz，H-10)，6.65(1H，d，J＝10.0Hz，H-4)，5.46(1H，d，J＝10.0Hz，H-3)，5.39(1H，ddd，J＝10.7，3.9，1.4Hz，H-27)，5.08(1H，dd，J＝7.2，5.9Hz，H-32)，3.50(1H，m，H-11)，3.49(1H，m，H1-26)，3.38(1H，dd，J＝10.5，1.9Hz，H-37)，3.28(1H，br dd，J＝15.1，5.6Hz，H1-31)，3.23(1H，dd，J＝15.0，7.4Hz，H2-31)，2.83(1H，ddd，J＝15.8，3.9，2.4Hz，H2-26)，2.50(1H，d，J＝9.4Hz，H-22)，2.34(1H，dd，J＝13.5，4.7Hz，H1-21)，2.05(1H，m，H1-20)，1.71(3H，s，H-34)，1.68(1H，m，H1-36)，1.64(3H，s，H-35)，1.63(3H，s，H-25)，1.62(1H，m，H2-20)，1.58(3H，s，H-29)，1.52(1H，m，H2-36)，1.44(3H，s，H-19)，1.37(1H，dd，J＝13.5，9.5Hz，H2-21)，1.27(3H，s，H-24)，1.18(6H，s，H-39、H-40)。

13C-NMR(150MHz，CDCl3)：δ203.26(C-12)，179.32(C-8)，168.56(C-30)，160.44(C-18)，157.89(C-16)，157.53(C-6)，135.74(C-27)，134.69(C-10)，133.85(C-9)，131.67(C-33)，129.00(C-28)，125.28(C-3)，122.40(C-32)，116.13(C-4)，108.14(C-17)，102.95(C-5)，100.68(C-7)，90.47(C-14)，84.20(C-13)，83.57(C-23)，80.56(C-2)，78.09(C-37)，73.63(C-38)，49.07(C-22)，47.05(C-11)，36.88(C-20)，29.99(C-26)，29.86(C-25)，29.11(C-24)，26.39(C-19)，25.89(C-39)，25.66(C-35)，25.56(C-36)，25.22(C-21)，23.40(C-40)，21.76(C-31)， 20.81(C-29)，18.07(C-34)。

The EIMS data presentation of product Gh-3271: have a part peak [M]+position in m/z 662 and a benchmark peak position at m/z 545 (100), and HREIMS data presentation [M]+m/z 662.3097, this represents that the molecular formula of product Gh-3271 is C38H46O10, more than gambogic acid 34 mass units.

1H-, 13C-NMR and HMQC spectrum display of product Gh-3271: the side chain be connected on C-2 position is different from gambogic acid institute tool person, and 3,4-dihydroxyl-4-methyl amyl instead of the 4-methyl-3-pentenyl of gambogic acid.In addition, 1H-NMR and the 13C-NMR spectrum display of product Gh-3271: have 1 primary hydroxyl groups [δ 3.38 (1H, dd, J=10.5,1.9Hz); δ 78.09] and 1 containing oxygen level Four carbon (δ 73.63).

The 1H-1H COSY spectroscopic data display of product Gh-3271: hydroxyl methine protons (hydroxymethine proton) [δ 3.38 (1H, dd, J=10.5,1.9Hz)] and δ 1.52 (1H, m) are coupled; δ 1.52 (1H, m) and δ 2.05 (1H, m, H1-20) are coupled; Two adjacent methene proton [δ 2.05 (H1-20), δ 1.62 (H2-20); δ 1.68 (H1-36), δ 1.52 (H2-36)] be coupled mutually.This proves that hydroxyl methine protons is H-37.

Confirm from the HMQC spectroscopic data of product Gh-3271: δ 36.88 (C-20), δ 25.56 (C-36) and δ 73.63 (C-38).This proves that primary hydroxyl groups is positioned at C-37, and three grades of oh groups are positioned at C-38.

Find from the HMBC spectroscopic data of product Gh-3271: δ 3.38 (H-37) is associated with δ 36.88 (C-20); δ 2.05 (H1-20) and δ 1.62 (H2-20) is all associated with δ 25.56 (C-36), δ 78.09 (C-37), δ 80.56 (C-2), δ 125.28 (C-3) and δ 26.39 (C-19); δ 5.46 (H-3) is associated with δ 80.56 (C-2), δ 26.39 (C-19) and δ 102.95 (C-5); δ 1.44 (H-19) is associated with δ 80.56 (C-2), δ 125.28 (C-3), δ 116.13 (C-4) and δ 160.44 (C-18); δ 1.18 (H-39, H-40) is associated with δ 78.09 (C-37) and δ 73.63 (C-38).This proves that the side chain be connected on C-2 position is 3,4-dihydroxyl-4-methyl amyl.

Find from the NOESY spectroscopic data of product Gh-3271: δ 7.50 (H-10) is associated with δ 3.50 (H-11); δ 3.50 (H-11) is associated with δ 2.34 (H1-21), δ 1.37 (H2-21); δ 1.37 (H2-21) is associated with δ 2.50 (H-22) and δ 1.27 (H-24); δ 2.50 (H-22) is associated with δ 1.63 (H-25); δ 5.39 (H-27) is associated with δ 1.58 (H-29).This proves that the three-dimensional arrangement of this part is identical with gambogic acid institute tool person, is all 11S, 13R, 14S, 22S configuration, and H-27 and carboxylic group (C-30) are trans (trans), and double bond Δ 27,28 is Z configuration.In addition, because δ 3.38 (H-37) is associated with δ 1.18 (H-39, H-40), and δ 2.05 (H1-20) is associated with δ 1.58 (H-29), so C-2 is S configuration.

Comprehensive above data, product Gh-3271 is identified is a compounds with the following chemical structure formula:

Product Gh-3271 is named as epiformoxanthone J (epiformoxanthone J), and { IUPAC names: 2-butylene acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydrochysene-8-hydroxyl-3, 3, 11-trimethylammonium-13-(3-methyl-2-butene base)-11-(3, 4-dihydroxyl-4-methyl amyl)-7, 15-dioxy-1, 5-methylene radical-1H, 3H, 11H-furo [3.4-g] piperazine is muttered also [3.2-b]-1-, base]-, (2Z)-[2-butenoic acid, 2-methyl-4-[(1R, 3aS, 5S, 11S, 14aS)-3a, 4, 5, 7-tetrahydro-8-hydroxy-3, 3, 11-trimethyl-13-(3-methyl-2-butenyl)-11-(3, 4-dihydroxy-4-methylpentyl)-7, 15-dioxo-1, 5-methano-1H, 3H, 11H-furo [3.4-g] pyrano [3.2-b] xanthen-1-yl]-, (2Z)-].

Conclusion:

The experimental result that physics and chemistry analysis obtain is carried out according to the product above for 35 provenance self-separation parts 1 to 3, in these 35 kinds of products, there are 18 kinds to be novel compound, have 17 kinds to be known compound (see table 4 below).In addition, in vitro anticancer test due to known product Gh-47, Gh-631, Gh-4601, Gh-4602 and Gh-2301 has been disclosed in this Taiwan Patent case TW I282280, therefore applicant's pharmacological evaluation of selecting other the 30 kinds of products except these 5 kinds of products to carry out below.

The product of table 4.35 provenance self-separation part 1 to 3

Embodiment 4. preparation is derived from the purified separate part TSB-14A of the acetone extract product TSB-14 of Gamboge to separate part TSB-14E

The acetone extract product TSB-14 of the Gamboge of 3g is dissolved in the acetone/acetonitrile (v/v=1: 9) of 30mL, and according to above " general operation procedure " when described in method carry out half preparation scale RP-HPLC, wherein movement is mixing solutions/95% acetonitrile of 0.0475%TFA and 5% acetonitrile mutually, and the gradient elution of mobile phase (gradient elution) is carried out in mode below: in 100 minutes, the mixing solutions of TFA and acetonitrile fades to 32% by 40%, and acetonitrile fades to 68% by 60%.The half preparation scale RP-HPLC elution figure obtained is shown in Fig. 6.

B, roughing out part TSB-14A are to the preparation of roughing out part TSB-14E:

Half preparation scale RP-HPLC elution figure (see Fig. 6) that A item obtains above is divided into 5 sections (sections) (namely section A is to section E) by applicant, and wherein the residence time of section A is the 0th to the 13.7th minute; The residence time of section B is the 13.7th to the 20.4th minute; The residence time of section C is the 20.4th to the 26.1st minute; The residence time of section D is the 26.1st to the 34.9th minute; And the residence time of section E is the 34.9th to the 53.8th minute.

The acetone extract product TSB-14 of the Gamboge of 3g is dissolved in the acetone/acetonitrile (v/v=1: 9) of 30mL, and according to above " general operation procedure " when described in method carry out half preparation scale RP-HPLC, wherein movement is mixing solutions/95% acetonitrile of 0.0475%TFA and 5% acetonitrile mutually, and the gradient elution of mobile phase (gradient elution) is carried out in mode below: in 100 minutes, the mixing solutions of TFA and acetonitrile fades to 32% by 40%, and acetonitrile fades to 68% by 60%.When gradient elution, collect the washings A to E corresponding to section A to section E respectively according to Fig. 6.

In addition, in order to obtain a large amount of washings A to E, half above-mentioned preparation scale RP-HPLC50 time is repeated.Obtained washings A to E is collected in the amber glass bottle of 4L respectively, then carries out concentrated with vacuum revolution thickener (vacuum rotatory evaporator) and obtain enriched material A to E.Then, enriched material A to E carries out distribution with H2O with ethyl acetate (ethyl acetate) respectively and is separated (partitioning), then cleans organic layer to remove TFA with H2O, then gives drying with anhydrous Na 2SO4.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener, and obtain roughing out part TSB-14A, TSB-14B, TSB-14C, TSB-14D and TSB-14E.

C, roughing out part TSB-14A are to the purifying of roughing out part TSB-14E:

In order to purification of crude separate part TSB-14A is to roughing out part TSB-14E, roughing out part TSB-14A obtained in the middle of B item is above dissolved in the acetone of 10mL respectively to roughing out part TSB-14E, and according to above " general operation procedure " when described in method carry out half preparation scale RP-HPLC, wherein movement is the 0.05%TFA aqueous solution/95% acetonitrile mutually, and about the gradient elution of mobile phase be carry out according to the condition below shown in table 5.

Table 5. is used in the mobile and gradient elution condition in half preparation scale RP-HPLC

Roughing out part	Operational condition
		TSB-14A	In 80 minutes, the TFA aqueous solution fades to 40% by 58%, and acetonitrile fades to by 42%

	60％。
		tSB-14B	in 80 minutes, the TFA aqueous solution fades to 40% by 70%, and acetonitrile fades to 60% by 30%.
tSB-14C	in 80 minutes, the TFA aqueous solution fades to 40% by 56%, and acetonitrile fades to 60% by 44%.
		tSB-14D	in 80 minutes, the TFA aqueous solution fades to 40% by 52%, and acetonitrile fades to 60% by 48%.
tSB-14E	in 80 minutes, the TFA aqueous solution fades to 40% by 48%, and acetonitrile fades to 60% by 52%.

When gradient elution, collect the washings of the roughing out part TSB-14A corresponding to each section to roughing out part TSB-14E according to the section A to section E (see Fig. 6) of the half preparation scale RP-HPLC elution figure obtained in the middle of A item above.Each washings carries out distribution with H2O with ethyl acetate respectively and is separated, and then, cleans organic layer to remove TFA, then give drying with anhydrous Na 2SO4 with H2O.After filtration, remove the organic solvent in filtrate with vacuum revolution thickener, and obtain separate part TSB-14A, TSB-14B, TSB-14C, TSB-14D and the TSB-14E through preliminary purification.

Separate part TSB-14A, TSB-14B, TSB-14C, TSB-14D and TSB-14E through preliminary purification is repeated half above-mentioned preparation scale RP-HPLC to carry out further purifying, until half preparation scale RP-HPLC elution figure of each separate part all presents consistent situation.After completing purifying, purified separate part TSB-14A (125mg), TSB-14B (85mg), TSB-14C (52mg), TSB-14D (267mg) and TSB-14E (1026mg) can be obtained.

The characterized of D, purified separate part TSB-14A to separate part TSB-14E:

The purified separate part TSB-14A to TSB-14E (respectively getting 100mg) obtained in the middle of C item is above dissolved in the acetone of 1mL respectively, and according to C item above when described in method carry out half preparation scale RP-HPLC, and obtain half preparation scale RP-HPLC elution figure (see Fig. 7 to Figure 11) of purified separate part TSB-14A to TSB-14E.Can know after Fig. 7 to Figure 11 is done a comparison with Fig. 1 of embodiment 1 above respectively and find out, the half preparation scale RP-HPLC elution figure of purified separate part TSB-14A to TSB-14E is respectively containing crest 1 to 12, crest 13 to 15, crest 16 to 20, crest 21 to 24 and crest 25 to 35.This represents that purified separate part TSB-14A contains product Gh-3261, Gh-3271, Gh-3272, Gh-3311, Gh-3332, Gh-1036, Gh-3291, Gh-631, Gh-1052, Gh-3351, Gh-3353 and Gh-3352; Purified separate part TSB-14B contains product Gh-47, Gh-4602 and Gh-4601; Purified separate part TSB-14C contains product Gh-1601-A, Gh-1050, Gh-1602, Gh-1631 and Gh-2641-1; Purified separate part TSB-14D contains product Gh-2501, Gh-2642, Gh-2507 and Gh-2505; And purified separate part TSB-14E contains product Gh-2508, Gh-2603-1, Gh-2603-2, Gh-1641, Gh-1642, Gh-2605, Gh-2606, Gh-2607-B, Gh-2607-1A, Gh-2301 and Gh-4301.

The pharmacological evaluation (Pharmacological experiments of acetone-extracted product TSB-14ofgamboge resin) of the acetone extract product TSB-14 of embodiment 5. Gamboge

In order to inquire into the acetone extract product TSB-14 of the Gamboge disclosed in this Taiwan Patent case TW I282280 except having the ability of inhibition tumor cell growth, whether also have other biological activity, pharmacological testing is below carried out.

Pharmacological testing 1. includes the activity of formula for cancer-associated proteins matter (cancer-related proteins) or the impact of binding ability of the acetone extract product TSB-14 of Gamboge

Be that the ratio of 9: 1 (wt/wt) mixes by the acetone extract product TSB-14 obtained by the embodiment 1 according to this Taiwan Patent case TW I282280 and brown sugar with one, and obtain the TSB-9 that fills a prescription.About this formula TSB-9 for the activity of cancer-associated proteins matter or the impact of binding ability be committee by MDSPharma Services on behalf of carrying out, wherein these tests are with reference to D.Riendeau et al. (1997), Can.J.Physiol.Pharmacol., 75:1088-1095, T.D.Warner etal. (1999), Proc.Natl.Acad.Sci.USA, 96:7563-7568, C.Ditchfiledet al. (2003), J.Cell Biol., 161:267-280, N.Gray et al. (1999), Curr.Med.Chem., 6:859-875, K.Cheng and J.G.Koland (1996), J.Biol.Chem., 271:311-318, K.Farley et al. (1992), Anal.Biochem., 203:151-157, E.Buchdunger et al. (1996), Cancer Res., 56:100-104, D.Q.Guo et al. (2000), J.Biol.Chem., 275:11216-11221, E.G.Barbacciet al. (2003), Cancer Res., 63:4450-4459, E.Liu et al. (1992), Oncogene, 7:1027-1032, T.Ozawa et al. (1998), Anal.Chem., 70:2345-2352 and J.D.Obourn et al. (1993), Biochem., 32:6229-6236 when described in method and slightly modified.

In this experiment, the experimental concentration of formula TSB-9 is set to 10 μ g/mL, and use 9 kinds of cancer-associated proteins matter { to comprise ring oxygenation-2 (cyclooxygenase-2, COX-2), protein thread amino acid/Soviet Union's amine acid kinase (protein serine/threonine kinase) AURKB (Aurora-B kinases) and CDC2/CCNB1 [cdk1/ cell periodic protein B (cyclin B)], protein tyros kinase (protein tyrosine kinase) ABL1 (ABL), EGF-R ELISA (Epidermal Growth Factor Receptor, EGFR), ERBB2 (HER2), insulin receptor (insulin receptor) and KDR (VEGFR-2), and estrogen receptor alpha (estrogenreceptor α, ER α) } test, and be shown in table 6 below about the effect of each test substances and the cancer types corresponding to it.

Table 6. is used to the cancer-associated proteins matter carrying out testing

Test substances

Effect

The cancer types relevant with test substances

Ring oxygenation-2	Cause inflammation effect, promote that tumor growth and inhibition tumor cell carving die (apoptosis)	Colorectal carcinoma (colorectal cancer) and cancer of the stomach (gastric cancer)
			Protein thread amino acid/Soviet Union's amine acid kinase, AURKB (Aurora-B kinases)	Regulating cell divides, and height indicator can cause numerical abnormalities of chromosomes now, promotes that cancer generates	Nonsmall-cell lung cancer (non-small cell lung cancer), the cancer of the brain (brain cancer) and thyroid carcinoma (thyroid cancer)
Protein thread amino acid/Soviet Union's amine acid kinase, CDC2/CCNB1 (cdk1/ cell periodic protein B)	Regulating cell divides, and height indicator can make cell fission out of hand now, promotes that cancer generates	Nasopharyngeal carcinoma (nasopharyngral cancer)
			Protein tyros kinase, ABL1 (ABL)	Promote growth of cancer cells	Chronic lymphocytic leukemia (chronic myelocytic leukemia) and T-cell acute lymphoid blast cell leukemia (T-cell acute lymphoblastic leukemia)
Protein tyros kinase, EGF-R ELISA	Activation signals pipeline (signal pathway), promotes hyperplasia	Lung cancer (lung cancer) and colorectal carcinoma (colon cancer)
			Protein tyros kinase, ERBB2 (HER2)	Activation signals pipeline, promotes hyperplasia	Breast cancer (breast cancer) and leukemia (leukemia)
Protein tyros kinase, insulin receptor	Meeting activation signals pipeline after being combined with Regular Insulin, promotes that hyperplasia and T suppression cell carving die	Breast cancer, colorectal carcinoma, lung cancer, liver cancer (liver cancer) and ovarian cancer (ovarial cancer)
			Protein tyros kinase, KDR (VEGFR-2)	Promote vasculogenesis (angiogenesis) and tumor growth	Breast cancer, lung cancer, liver cancer, renal cancer (renal cancer) and cancer of pancreas (pancreas cancer)
Estrogen receptor alpha	Promote hyperplasia	Breast cancer, lung cancer and carcinoma of endometrium (endometrial cancer)

According to the laboratory report of MDS Pharma Services, formula TSB-9 is shown in table 7 below for the activity of above-mentioned 9 kinds of cancer-associated proteins matter or the inhibiting rate (%) of binding ability.

Table 7. is filled a prescription the activity of TSB-9 for cancer-associated proteins matter or the inhibiting rate of binding ability

Test substances	Inhibiting rate (%)
		Ring oxygenation-2	83
Protein thread amino acid/Soviet Union's amine acid kinase, (Aurora-B swashs AURKB	96

Enzyme)
		Protein thread amino acid/Soviet Union's amine acid kinase, CDC2/CCNB1 (cdk1/ cell periodic protein B)	94
Protein tyros kinase, ABL1 (ABL)	99
		Protein tyros kinase, EGF-R ELISA	94
Protein tyros kinase, ERBB2 (HER2)	86
		Protein tyros kinase, insulin receptor	100
Protein tyros kinase, KDR (VEGFR-2)	99
		Estrogen receptor	66

From result shown in table 7, the acetone extract product TSB-14 of Gamboge has the effectiveness suppressing cancer-associated proteins matter activity or binding ability significantly.Because these cancer-associated proteins matter are relevant with kinds of tumors/cancer (such as colorectal carcinoma, cancer of the stomach, nonsmall-cell lung cancer, the cancer of the brain, thyroid carcinoma, nasopharyngeal carcinoma, chronic lymphocytic leukemia, T-cell acute lymphoid blast cell leukemia, lung cancer, colorectal carcinoma, breast cancer, leukemia, liver cancer, ovarian cancer, renal cancer, cancer of pancreas and carcinoma of endometrium), applicant is inference accordingly: the acetone extract product TSB-14 of Gamboge is available for treatment lesion/cancer disease.

Pharmacological testing 2. is analgesic test (in vivo analgesia test) in vivo

Be that the ratio of 9: 1 (wt/wt) mixes by the acetone extract product TSB-14 obtained by the embodiment 1 according to this Taiwan Patent case TW I282280 and brown sugar with one, then ground and obtained there is the formula TSB-9-W that median size (particle size) is about 10 μm.The formula TSB-9-W obtained entrusts by MDS Pharma Services on behalf of the in vivo analgesic test carried out below.

Male mice derivative for CD-1 (Crl.) is divided into 3 groups (respectively organizing n=5) randomly, comprising 1 experimental group (namely TSB-9-W group) and 2 control groups [namely supporting agent Normal group (vehicle normal control) and morphine positive control group (morphine positivecontrol)].The mouse of TSB-9-W group by oral administration medicine supplying with the TSB-9-W that fills a prescription (dosage is for 100mg/kg), and the mouse of supporting agent Normal group and morphine positive control group respectively by oral administration medicine supplying with 2%Tween 80 (dosage is for 10mL/kg) and morphine (dosage is for 30mg/kg).The 1st hour after oral administration medicine supplying, by 2% Formalin solution (formalin solution) sole of the foot hemostasis (subplantar injection) of 0.02mL to the hind paw of each group of mouse, and during after injection the 0th to the 5th minute and the hind paw noting down mouse during the 15th to the 30th minute lick and lick number of times (hind paw licking time).Hind paw about each group of mouse is licked and licked inhibiting rate (%) is be calculated by following formula (1):

Formula (1): A=(B-C)/B × 100

Wherein: A=hind paw is licked and licked inhibiting rate (%)

The hind paw of B=supporting agent Normal group mouse licks the mean value of licking number of times

The hind paw that C=respectively organizes mouse licks the mean value of licking number of times

If the hind paw obtained lick lick inhibiting rate be >=50%, this indicates the analgesic activity shown.

The experimental result obtained is shown in table 8 below.From result shown in table 8, with supporting agent Normal group mouse in comparison, by oral administration medicine supplying with during the mouse of the TSB-9-W that fills a prescription after Formalin injection of solution the 0th to the 5th minute and the 15th to the 30th point of bell, their hind paw is licked and is licked inhibiting rate and be enhanced significantly, especially, during the 0th to the 5th minute, hind paw is licked and is licked inhibiting rate and can reach 68%.This experimental result shows: the acetone extract product TSB-14 of Gamboge can reach the effectiveness of inhibition of pain.

The table 8. respectively hind paw organized measured by mouse is licked and is licked inhibiting rate

*: when hind paw lick lick inhibiting rate be >=50%, indicate the analgesic activity shown.

Pharmacological testing 3. is anti-inflammation test (in vivo anti-inflammation test) in vivo

Be that the ratio of 9: 1 (wt/wt) mixes by the acetone extract product TSB-14 obtained by the embodiment 1 according to this Taiwan Patent case TW I282280 and brown sugar with one, then ground and obtained there is the formula TSB-9-W1 that median size is about 5 μm.The formula TSB-9-W1 obtained is that committee is tested on behalf of the in vivo anti-inflammation carried out below by MDSPharma Services.

Male Wistar rat (body weight is about 160 ± 5g) is divided into 4 groups (respectively organizing n=5) randomly, comprising 2 experimental group (namely TSB-9-W1-1 group and TSB-9-W1-2 group) and 2 control groups [namely supporting agent Normal group and aspirin positive control group (aspirin positivecontrol)].After fasting is overnight, the rat of TSB-9-W1-1 group and TSB-9-W1-2 group is the formula TSB-9-W1 of 10mg/kg and 30mg/kg respectively with dosage by oral administration medicine supplying, and the rat of supporting agent Normal group and aspirin positive control group respectively by oral administration medicine supplying with 2%Tween 80 (dosage is for 10mL/kg) and aspirin (dosage is for 150mg/kg).The 1st hour after oral administration medicine supplying, by injection (intraplantar injection) in 1% carrageeman suspension (carrageenan suspension) sole of the foot of 0.1mL to the right back sole of each group of rat, and after injection the 3rd hour, (diameter is 25mm to use one to have water unit (water cell); #7157, UGO Basile, Italy) volume determining instrument (plethysmometer) (#7150, UGO Basile, Italy) measure the left back sole of rat and the volume of right back sole.The hind paw edema volume of rat represents with the difference of person measured by the volume measured by Rat Right hind paw and left back sole.Hind paw oedema inhibiting rate (%) about each group of rat is calculated by following formula (2):

Formula (2): D=(E-F)/E × 100

Wherein: D=hind paw oedema inhibiting rate (%)

The mean value of the hind paw edema volume of E=supporting agent rats in normal control group

F=respectively organizes the mean value of the hind paw edema volume of rat

If the hind paw oedema inhibiting rate obtained is >=30%, and this indicates that the anti-inflammation shown is active.

The experimental result obtained is shown in table 9 below.From result shown in table 9, with supporting agent Normal group mouse in comparison, the 3rd hour after being injected in the sole of the foot of 1% carrageeman suspension with the rat of the TSB-9-W1 that fills a prescription by oral administration medicine supplying, their hind paw oedema inhibiting rate is enhanced significantly, especially, under being the dosage of 30mg/kg one, hind paw oedema inhibiting rate can be made to reach to 30%.This experimental result shows: the acetone extract product TSB-14 of Gamboge reaches the effectiveness to anti-inflammation in the mode of dose-dependency (dosage-dependent).

The hind paw oedema inhibiting rate measured by rat respectively organized by table 9.

Group	Hind paw oedema inhibiting rate (%)
		Supporting agent Normal group	0
TSB-9-W1-1 group	17
		TSB-9-W1-2 group	30*
Aspirin positive control group	40*

*: when hind paw oedema inhibiting rate be >=30%, indicate that the anti-inflammation that shows is active.

Embodiment 6. is derived from the separate part of acetone extract product TSB-14 and the pharmacological evaluation of purified compound of Gamboge

In order to inquire into according to separate part TSB-14A of the present invention to separate part TSB-14E and by according to separate part 1 to separate part 3 of the present invention by the biological activity of 30 kinds of products (not comprising Gh-47, Gh-631, Gh-4601, Gh-4602 and Gh-2301) be further purified out, pharmacologically active analysis is below carried out.

Pharmacological testing 1. is anticancer test (in vitro anti-cancer test) in vitro

About according to separate part TSB-14A of the present invention to separate part TSB-14E and by according to separate part 1 to separate part 3 of the present invention by the in vitro anticancer test of the 30 kinds of products (not comprising Gh-47, Gh-631, Gh-4601, Gh-4602 and Gh-2301) be further purified be committee by MDSPharma Services on behalf of carrying out.

This anticancer test is mainly used for the proliferative effect of detecting one drug candidates (drug candidate) for cancer cells, central involved action principle is that viable cell can by alamarBlue reagent (AbDSerotec, UK) by the state of oxidation (the non-fluorescent originally without fluorescent, blue) the reduction form (fluorescent becoming tool fluorescent is reacted through metabolism, red) ability, and according to the measured fluorescent data results produced by alamarBlue reagent, the propagation situation of viable cell and cytoactive can be quantized and detect.

In this experiment, the experimental concentration of drug candidates is set to 0.01,0.1,1,10 and 100 μ g/mL, and uses 6 kinds of human cancer cell's strains and a kind of normal human cell's strain (see table 10) coming to carry out.In addition, use the diformazan Asia (dimethylsulfoxide, DMSO) of 40% as Normal group (normal control) and use mitomycin (mitomycin) as positive control group (positive control).

Table 10. is used to the cell strain carrying out in vitro anticancer test

Cell strain title	Source
		People Class breast cancer cell MCF7 (human breast adenocarcinoma cell MCF7)	ATCC HTB-22
Human colon's adenocarcinoma cell HT-29 (human colon adenocarcinoma cell HT-29)	ATCC HTB-38
		Myelogenous leukemia cells HL-60 (human promyelocytic leukemia cell HL-60) before the mankind	ATCC CCL-240
Human hepatocytes cancer cells HepG2 (human hepatocel lular carcinoma cell HepG2)	ATCC HB-8065
		Human Lung Cancer cell A549 (human lung carcinoma cell A549)	ATCC CCL-185
Human tissue cells's property lymphoma cell U937 (human histocytic lymphoma cell U937)	ATCC CRL-1593
		Human umbilical vein's endotheliocyte (human umbilical venal epithelial cell, HUVEC)	ATCC CRL-1730

According to the laboratory report of MDS Pharma Services, according to separate part TSB-14A of the present invention to separate part TSB-14E and by according to separate part 1 to separate part 3 of the present invention purified go out 30 kinds of products for 50% inhibition concentration (the 50% inhibition concentration of above-mentioned 6 kinds of human cancer cell's strains and a kind of normal human cell's strain, I C50) and 50% lethal concentration (50%lethal concentration, LC50) be shown in respectively in table 11 below to table 14.

The separate part TSB-14A of the acetone extract product of table 11. Gamboge is to being separated

Part TSB-14E is for 50% inhibition concentration (IC50 of cancer cells; μ g/mL)

Mitomycin (μM)

0.033

0.15

0.16

0.042

0.23

0.56

The separate part TSB-14A to separate part TSB-14E of the acetone extract product of table 12. Gamboge is for 50% lethal concentration (LC50 of cancer cells; μ g/mL)

30 kinds of products that table 13. self-separation part 1 to separate part 3 is purified into are for 50% inhibition concentration (IC50 of cancer cells; μ g/mL)

Mitomycin (μM)	0.15	0.49	0.098	0.11	0.099	0.072	0.12
								Gh-1641	1.7	1.4	8.0	0.86	0.47	9.4	0.18
Gh-1642	2.4	1.9	6.0	1.1	0.51	8.8	0.18
								Mitomycin (μM)	0.080	0.51	0.062	0.075	0.15	0.098	0.10
Gh-2642	3.0	12	9.0	11	1.5	1.3	2.4
								Mitomycin (μM)	0.13	0.57	0.12	0.062	0.24	0.17	0.13
Gh-1602	0.27	0.94	1.0	1.3	0.40	0.14	0.25
								Mitomycin (μM)	0.079	0.37	0.20	0.073	0.21	0.19	0.088
Gh-2607-B	0.10	0.30	0.13	0.31	0.10	0.10	0.07
								Gh-2607-1A	0.12	0.24	0.49	0.43	0.19	0.15	0.43
Gh-1601-A	0.72	0.17	0.56	1.3	0.11	0.09	0.19
								Gh-3352	0.075	0.18	0.26	0.16	0.18	0.11	0.084
Mitomycin (μM)	0.15	0.11	0.24	0.21	0.098	0.18	0.15

30 kinds of products that table 14. self-separation part 1 to separate part 3 is purified into are for 50% lethal concentration (LC50 of cancer cells; μ g/mL)

Gh-2607-1A
								Mitomycin (μM)	1.8	9.3	1.3	1.3	1.9	2.0	6.6
Gh-1641	33	4.4	32	4.5	3.0	36	1.3
								Gh-1642	29	4.8	35	5.3	2.8	33	1.3
Mitomycin (μM)	5.8	8.5	1.2	2.3	3.6	1.9	2.4
								Gh-2642	10	42	32	29	4.6	4.1	11
Mitomycin (μM)	＞10	＞10	2.3	1.6	4.5	0.87	2.4
								Gh-1602	4.0	4.2	3.3	3.5	3.7	0.69	0.94
Mitomycin (μM)	＞10	＞10	1.8	2.4	5.3	0.82	1.7
								Gh-2607-B	0.68	1.0	3.1	4.0	0.33	0.25	1.6
Gh-2607-1A	1.5	0.84	2.6	3.9	0.45	0.37	1.5
								Gh-1601-A	4.4	0.77	3.0	4.1	6.6	0.28	5.4
GH-3352	1.8	0.68	0.93	2.0	0.99	0.26	0.85
								Mitomycin (μM)	17	22	0.89	5.2	4.4	0.85	1.8

Result shown in from table 11 to table 14, according to separate part TSB-14A of the present invention to separate part TSB-14E and by according to separate part 1 to separate part 3 of the present invention purified go out 18 kinds of compounds there is Tumor suppression/growth of cancer cells all significantly and kill the effectiveness of lesion/cancer cell.Especially, the acetone extract product TSB-14 institute tool person of Gamboge that discloses in TW I282280 than applicant haply of the effectiveness of these separate parts TSB-14A to separate part TSB-14E on Tumor suppression/growth of cancer cells is for good.Therefore, according to separate part TSB-14A of the present invention to separate part TSB-14E and by according to separate part 1 to separate part 3 of the present invention purified go out 18 kinds of compounds all there are high potentiality as a cancer therapy drug.

Pharmacological testing 2. is derived from the activity of separate part TSB-14A to separate part TSB-14E for cancer-associated proteins matter or the impact of binding ability of the acetone extract product of Gamboge

About according to separate part TSB-14A of the present invention to separate part TSB-14E for the activity of cancer-associated proteins matter or the impact of binding ability be equally committee by MDS Pharma Services on behalf of carrying out, acquired results is shown in table 15 below.

The activity of separate part TSB-14A to separate part TSB-14E for cancer-associated proteins matter of the acetone extract product of table 15. Gamboge or the inhibiting rate (%) of binding ability

TSB-14A

TSB-14B

TSB-14C

TSB-14D

TSB-14E

Ring oxygenation-2	35	74	99	77	83
						Protein thread amino acid/Soviet Union's amine acid kinase, AURKB (Aurora-B kinases)	60	78	86	85	90
Protein thread amino acid/Soviet Union's amine acid kinase, CDC2/CCNB1 (cdk1/ cell periodic protein B)	2	69	80	67	46
						Protein tyros kinase, ABL1 (ABL)	97	104	105	102	102
Protein tyros kinase, EGF-R ELISA	79	90	96	82	85
						Protein tyros kinase, ERBB2 (HER2)	78	84	84	84	64
Protein tyros kinase, insulin receptor	13	81	78	66	46
						Protein tyros kinase, KDR (VEGFR-2)	-12	10	44	37	65
Estrogen receptor	45	67	83	89	75

From result shown in table 15, have all significantly according to separate part TSB-14A of the present invention to separate part TSB-14E and suppress the activity of cancer-associated proteins matter or the effectiveness of binding ability.Because these cancer-associated proteins matter are relevant with kinds of tumors/cancer (such as colorectal carcinoma, cancer of the stomach, nonsmall-cell lung cancer, the cancer of the brain, thyroid carcinoma, nasopharyngeal carcinoma, chronic myelognous chronic myeloid leukemia, T-cell acute lymphoid blast cell leukemia, lung cancer, colorectal carcinoma, breast cancer, leukemia, liver cancer, ovarian cancer, renal cancer, cancer of pancreas and carcinoma of endometrium), applicant is inference accordingly: be available for treatment lesion/cancer disease according to separate part TSB-14A of the present invention to separate part TSB-14E.

Pharmacological testing 3. is derived from the in vivo analgesic test of separate part TSB-14A to separate part TSB-14E of the acetone extract product of Gamboge

The in vivo analgesic test of relevant separate part TSB-14A to separate part TSB-14E according to the present invention is entrusted by MDS Pharma Services on behalf of carrying out.

Male mice (body weight is about 22 ± 3g) derivative for CD-1 (Crl.) is divided into 7 groups (respectively organizing n=6) randomly, comprising 5 experimental group (namely TSB-14A group, TSB-14B group, TSB-14C group, TSB-14D group and TSB-14E group) and 2 control groups (namely supporting agent Normal group and morphine positive control group).About the working method of in vivo analgesic test be substantially with reference to embodiment 5 above " pharmacological testing 2. in vivo analgesic test " when described in method carry out, difference is: the mouse of TSB-14A to TSB-14E group respectively by oral administration medicine supplying with in the middle of the C item of embodiment 4 above the purified separate part TSB-14A to separate part TSB-14E (dosage is all 100mg/kg) that obtains, and the mouse of supporting agent Normal group by oral administration medicine supplying with 5%DMSO/2%Tween 80 (dosage is for 10mL/kg), and the after 2% Formalin injection of solution the 0th to the 5th minute, 10th to the 15th minute, 15th to the 20th minute, during 20th to the 25th minute and the 25th to the 30th minute, the hind paw of record mouse is licked and is licked number of times.

The experimental result obtained is shown in table 16 below.From result shown in table 16, with supporting agent Normal group mouse in comparison, respectively by oral administration medicine supplying with during after Formalin injection of solution the 0th to the 30th minute of the mouse of separate part TSB-14A of the present invention to separate part TSB-14E, their hind paw is licked and is licked inhibiting rate and be enhanced significantly.Especially, during whole experiment, to be licked with the hind paw of the mouse of separate part TSB-14E by oral administration medicine supplying and lick inhibiting rate and be maintained between 68 to 100%.This experimental result show: the effectiveness all to separate part TSB-14E according to separate part TSB-14A of the present invention with inhibition of pain, wherein with the analgesic effect of separate part TSB-14E for the best.

The table 16. respectively hind paw organized measured by mouse is licked and is licked inhibiting rate

The in vivo anti-inflammation that pharmacological testing 4. is derived from the separate part TSB-14A to separate part TSB-14E of the acetone extract product of Gamboge is tested

The in vivo anti-inflammation test of relevant separate part TSB-14A to separate part TSB-14E according to the present invention is entrusted by MDS Pharma Services on behalf of carrying out.

Male Wistar rat (body weight is about 155 ± 5g) is divided into 7 groups (respectively organizing n=5) randomly, comprising 5 experimental group (namely TSB-14A group, TSB-14B group, TSB-14C group, TSB-14D group and TSB-14E group) and 2 control groups (namely supporting agent Normal group and aspirin positive control group).About the working method of in vivo anti-inflammation test be substantially with reference to embodiment 5 above " pharmacological testing 3. in vivo anti-inflammation test " when described in method carry out, difference is: the rat of TSB-14A to TSB-14E group respectively by oral administration medicine supplying with in the middle of the C item of embodiment 4 above the purified separate part TSB-14A to separate part TSB-14E (dosage is all 30mg/kg) that obtains, and the rat of supporting agent Normal group by oral administration medicine supplying with 5%DMSO/2%Tween 80 (dosage is for 10mL/kg).

The experimental result obtained is shown in table 17 below.From result shown in table 17, with supporting agent Normal group mouse in comparison, the 3rd hour after being injected in the sole of the foot of 1% carrageeman suspension with the rat of separate part TSB-14A to TSB-14E of the present invention by oral administration medicine supplying respectively, their hind paw oedema inhibiting rate is enhanced significantly, especially, the hind paw oedema inhibiting rate of TSB-14A group, TSB-14C group and TSB-14E group rat all can reach to more than 30%.This experimental result show: the effectiveness all to separate part TSB-14E according to separate part TSB-14A of the present invention with anti-inflammation, wherein with the effect of separate part TSB-14E for the best.

The hind paw oedema inhibiting rate measured by rat respectively organized by table 17.

Group	Hind paw oedema inhibiting rate (%)
		Supporting agent Normal group	0
TSB-14A group	30*
		TSB-14B group	24
TSB-14C group	33*
		TSB-14D group	26
TSB-14E group	34*
		Aspirin positive control group	37*

The all documents and materials be quoted in this case specification sheets and patent document are merged in this case as reference data using their entirety.If when conflicting to some extent, the detailed description of this case will be got the upper hand (comprising in being defined in).

Although the present invention is described with reference to above-mentioned specific concrete example, a lot of modifications and variations can be made not deviating under scope and spirit of the present invention significantly.Therefore it is intended that the present invention is only by the restriction as examined attached claims those shown with literary composition.

The above, it is only preferred embodiment of the present invention, not any pro forma restriction is done to the present invention, although the present invention discloses as above with preferred embodiment, but and be not used to limit the present invention, any those skilled in the art, do not departing within the scope of technical solution of the present invention, make a little change when the technology contents of above-mentioned announcement can be utilized or be modified to the Equivalent embodiments of equivalent variations, in every case be do not depart from technical solution of the present invention content, according to any simple modification that technical spirit of the present invention is done above embodiment, equivalent variations and modification, all still belong in the scope of technical solution of the present invention.

Claims

1. a purifying is from Gamboge and be by compound selected among following formed group:

(1) a kind of compound with following shown chemical formula:

(2) a kind of compound with following shown chemical formula:

(3) a kind of compound with following shown chemical formula:

(4) a kind of compound with following shown chemical formula:

(5) a kind of compound with following shown chemical formula:

(6) a kind of compound with following shown chemical formula:

(7) a kind of compound with following shown chemical formula:

(8) a kind of compound with following shown chemical formula:

(9) a kind of compound with following shown chemical formula:

(10) a kind of compound with following shown chemical formula:

(11) a kind of compound with following shown chemical formula:

(12) a kind of compound with following shown chemical formula:

2., for suppressing a pharmaceutical composition for the growth of one or more lesion/cancer cells, it is characterized in that:

Described pharmaceutical composition contains one or more compounds as claimed in claim 1.

3. pharmaceutical composition as claimed in claim 2, is characterized in that:

Described lesion/cancer cell is selected from by following formed group: myelogenous leukemia cells, human hepatocytes cancer cells, Human Lung Cancer cell and human tissue cells's property lymphoma cell before mankind mastopathy cell, human colon's adenocarcinoma cell, the mankind.

4. be used for the treatment of a pharmaceutical composition for one or more lesion/cancer diseases, it is characterized in that:

5. pharmaceutical composition as claimed in claim 4, is characterized in that:

Described lesion/cancer disease is selected from by following formed group: colorectal carcinoma, cancer of the stomach, lung cancer, the cancer of the brain, thyroid carcinoma, nasopharyngeal carcinoma, leukemia, ovarian cancer, renal cancer, cancer of pancreas and carcinoma of endometrium.

6. pharmaceutical composition as claimed in claim 5, is characterized in that:

Described leukemia is chronic lymphocytic leukemia or T-cell acute lymphoid blast cell leukemia.

7. pharmaceutical composition as claimed in claim 5, is characterized in that:

Described lung cancer is nonsmall-cell lung cancer.