CN106366177A - Thymopentin methyl ester conjugate and preparation method thereof - Google Patents
Thymopentin methyl ester conjugate and preparation method thereof Download PDFInfo
- Publication number
- CN106366177A CN106366177A CN201610806260.7A CN201610806260A CN106366177A CN 106366177 A CN106366177 A CN 106366177A CN 201610806260 A CN201610806260 A CN 201610806260A CN 106366177 A CN106366177 A CN 106366177A
- Authority
- CN
- China
- Prior art keywords
- solution
- preparation
- fpsdm
- stirring
- methyl ester
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/66—Thymopoietins
Abstract
The invention relates to a thymopentin methyl ester conjugate and a preparation method thereof. The preparation method specifically comprises the following steps: dissolving Z-FPSDM-Arg(Pbf)-Lys(BOC)-OH and 1-hydroxybenzotriazole (HOBt)/DCC/MMF in anhydrous trichloromethane, adding N,N-ethyldiisopropylamine and other specific solvents for reacting while stirring, thereby obtaining an active ester A; dripping the active ester into an alkaline solution of tripeptide derivatives, adding N,N-diisopropylamine and other specific solvents, thereby obtaining a coarse product of the thymopentin methyl ester conjugate; and finally, performing deprotection, reduced pressure distillation, washing and drying, thereby obtaining an accurately modified thymopentin methyl ester conjugate. The preparation method disclosed by the invention is fewer in byproducts and high in synthetic efficiency. The thymopentin methyl ester conjugate has the effects of maintaining the biological activity of TP-5, relieving or avoiding enzymolysis of TP-5 and further obviously prolonging the in-vivo metabolic half-life, and is extremely high in clinical application value.
Description
Technical field
The present invention relates to a kind of Thymopentin methyl ester conjugates and preparation method thereof, especially relate to a kind of fixed member amount
Precision architecture drug modification thing-Thymopentin methyl ester conjugates and preparation method thereof.And the method is suitable to large-scale production.
Background technology
Thymopentin (thymopetidu, tp-5) is in endogenous polypeptide thymopoietin (thymopoietin) activity
Five fragments of peptides of the heart, by 5 Amino acid profiles, have and thymopoietin identical physiological action, the immunologic function to body
There is dual regulation, be clinically important immunologic balance regulator.Tp-5 can be used for used as adjuvant drug for antitumor, hepatitis B,
Autoimmune disease, the immunologic hypofunction such as worn with age；Also can as adjuvant, with many commonly-used Drug combination with
Heighten the effect of a treatment, including interferon, hormone, analgesic, depressor, the medicine of cardiovascular disease, medicine for central nervous system etc..
Tp-5 extremely unstable in vivo, is easily degraded by protease and aminopeptidase, and presently mainly injection or enteric agents are given
Medicine, but its serum half-life is only 30 seconds, this shortcoming leads to tp-5 to maintain the time of normal blood drug level short in vivo, loses efficacy
Soon it is necessary to frequent drug administration, to maintain medicine effect, increases patient's treatment cost, increases economy and the mental burden of patient.For
The shortcoming solving tp-5 half-life short, numerous researchers develop the Thymopentin of chemical modification, and such as aminoacid sequence is adjusted
Whole with replace, end group chemistry modify with peptide bond replace, the coupling etc. of inverso analog and cyclic peptide synthesis and other conjugates, its
Middle conjugates have: lipoamino acid, thymosin α1, cell-penetrating peptide and polyvinyl alcohol etc..But carry out only for drug modification above
Improve, the shortcoming fundamentally not solving tp-5 half-life short, with the continuous expansion of Clinical practice amount, need badly a internal
The long lasting immune balance adjustment class medicine of good stability.
Medicinal polymer has larger molecular weight, and itself does not participate in internal biochemical reaction, has no toxic side effect, but can bring one
A little assosting effects.Gradually paid close attention in pharmaceuticals industry and applied.Wherein interlinked formation " poly by chemical bond and medicine
Thing-medicament coupling thing ".
As the specific type in medicinal polymer, it removes the basic demand meeting medicinal polymer, except having safety
Good, Stability Analysis of Structures, be not involved in internal biochemical reaction, have no toxic side effect and wait outward, also have the following characteristics that 1) contain functional group,
Can be connected with each other by forming chemical bond under certain condition with the special groups in drug molecular structure；2) physicochemical property energy
Complementary with medicine, some properties of active compound thing can be efficiently modified after forming polymer-medicament coupling thing；3) can be many to formed
Medicine in polymers-medicament coupling thing produces good shielding action, thus overcoming or reducing medicine in vivo in metabolic process
The shortcoming occurring.
And most of medicinal polymer, be inherently be polymerized by basic structural unit, molecular weight is in normal distribution
Mixture.The conjugates being formed by this kind of polymer and bioactive molecule, also will be uneven because of polymer molecules amount and structure
One and the conjugates obtained by leading to are mixture.This shortcoming, be polymer-medicament coupling thing research and development increased much hidden
Suffer from.
Therefore by larger to synthesized tp-5 derivant and molecular weight, containing functional group, have single molecular structure and
Functional type precision architecture trim (fpsdm) of molecular weight is connected and obtains corresponding conjugates, and exploitation has potentially good
Trim-tp-5 analog the conjugates of good resistance to enzymolysis ability and internal stability are significant, and clinical value is high.
Content of the invention
In order to overcome in tp-5 structure c- end carboxyl and phenolic hydroxyl group to be easy to ionizing in environment in vivo, to passing through cell membrane
Form obstacle, and the easy weakness such as enzymolysis, internal metabolism half-life short, the technical problem to be solved is to provide one kind
By way of chemical bond is connected with methyl ester tp-5 fixed point, designs, synthesized functional type precision architecture drug modification thing-thymus five
Peptide methyl ester conjugates (fpsdm-tp-5) and preparation method thereof.
The technical scheme is that a kind of preparation side of Thymopentin methyl ester conjugates
Method, comprises the following steps:
1) z-fpsdm-arg (pbf)-lys (boc)-oh and hobt/dcc/mmf is dissolved in anhydrous chloroform, while stirring
Mix side and add n, n- diisopropylethylamine is reacted, and obtains active ester a；
2) described active ester a is added drop-wise in the alkaline solution of tripeptide derivative, is simultaneously introduced n, n- diisopropylamine etc.
Specific solvent, is stirred together to and reacts completely, then adds chloroform, and carry out successively washing, be dried, vacuum distillation,
Obtain crude product Thymopentin methyl ester conjugates；
3) add the removing reagent of the chloroform soln of trifluoroacetic acid of 5%-20% in described crude product, stir together
Mix, then carry out vacuum distillation and pump trifluoroacetic acid and chloroform solvent, obtain residue, then described residue is entered successively
Row washs, is dried, that is, obtain described Thymopentin methyl ester conjugates.
The concrete synthetic route of described Thymopentin methyl ester conjugates is as follows:
The invention has the beneficial effects as follows: the present invention carries out esterification to the c- end carboxyl of tp-5 first, then using having
Functional type precision architecture drug modification thing fpsdm (the functional precision of larger and unimodal molecular weight
Structure drug modifier), by way of chemical bond is connected with methyl ester tp-5 fixed point, designs, synthesized functional type
Precision architecture drug modification thing-Thymopentin methyl ester conjugates (fpsdm-tp-5).By the participation of fpsdm and played right
Enzyme shielding action, tp-5 is effectively alleviated or avoided to the fpsdm-tp-5 of preparation, while keeping tp-5 biological activity, in body
Interior enzymolysis so as to the internal metabolism half-life significantly extend, thus the shortcoming effectively overcoming tp-5；This is due in tp-5 methyl ester
Carboxyl protected, can effectively overcome its easy ionizing, cell membrane pass through ability shortcoming.Additionally, adopting unimodal molecular weight
The fpsdm and methyl ester tp-5 fixed point fpsdm-tp-5 that is connected and synthesizes there is brand-new chemical constitution.
The medicinal polymer fpsdm that the present invention adopts has the characteristic being significantly better than common drug polymer: has fixation
Chemical constitution, single molecular weight, it obtains single compound fpsdm-tp- of molecular weight fixation with medicine after fixed point is connected
5 conjugates, it is to avoid traditional drugss polymer molecules amounts is fixing and medicinal polymer-medicament coupling thing that cause is mixture
And reduce drug effect.
The conjugates being prepared can make fpsdm shielding action be effectively played, be avoided that again because conjugates become
The many drawbacks brought for mixture are potential, clinical application is more extensive, better efficacy, may replace the new of tp-5
Immunologic balance regulator.
On the basis of technique scheme, the present invention can also do following improvement.
Further, in step 1) described in stirring time be 45～75 minutes.
Further, in step 2) in, described alkaline solution is dichloromethane solution；The time of described stirring is little for 7～9
When；Described washing cleaning mixture used is sequentially successively: first washs with hcl solution, with solution washing and then molten with naoh again
Liquid washs, and is finally washed with saturation nacl solution；Desiccant used by described drying is na2so4Desiccant；Described decompression
Under temperature be 20～35 DEG C.
Further, in step 3) in, the time of described stirring is 3～5 hours；Described washing cleaning mixture used is
et2o；Temperature under described decompression is 20～35 DEG C.
Further, the synthesis step of described z-fpsdm-arg (pbf)-lys (boc)-oh is:
(1) by z-fpsdm and 1- hydroxyl -1h-1,2,3- triazoles -4- carboxylic acid, ethyl ester (hoct) are dissolved in anhydrous methylene chloride
(dcm), in, add n while stirring, n- DIC (dic) is reacted, and obtains active ester b；
(2) described active ester b is added drop-wise in the alkaline solution of h-arg (pbf)-lys (boc)-ome, is simultaneously introduced n-
Diisopropylethylamine (diea), is stirred together to and reacts completely；Add dichloromethane, then carry out successively again washing, be dried,
Vacuum distillation, obtains z-fpsdm-arg (pbf)-ome；
(3) gained z-fpsdm-arg (pbf)-lys (boc)-ome is added in meoh, adds lioh water while stirring
Solution is to reacting completely, then carries out decompression and pump solvent, obtains residue, described residue is carried out successively wash, be dried, subtract
After pressure distillation, recrystallization, that is, synthesize described z-fpsdm-arg (pbf)-lys (boc)-oh.
The concrete synthetic route of described z-fpsdm-arg (pbf)-lys (boc)-oh is as follows:
Further, in step (1), the time of described stirring is 30～60 minutes.
Further, in step (2), described alkaline solution is dichloromethane solution；The time of described stirring is little for 5～7
When；Described washing cleaning mixture used is sequentially successively: first washs with hcl solution, with solution washing and then molten with naoh again
Liquid washs, and is finally washed with saturation nacl solution；Desiccant used by described drying is na2so4Desiccant；Described decompression
Under temperature be 20～35 DEG C.
Further, in step (3), the time of described stirring is 2～4 hours；Described reduce pressure twice under temperature be
20～35 DEG C；Described washing cleaning mixture used is followed successively by hcl solution and saturated aqueous common salt；Desiccant used by described drying is
na2so4Desiccant；Dichloromethane that the material of described recrystallization is, et2O and normal hexane, wherein said dichloromethane, et2o
Ratio of weight and number with normal hexane is 1:6:1.
Due to employing above technical scheme, make what the present invention possessed to have the beneficial effects that:
The Thymopentin methyl ester conjugates of present invention preparation, especially employ the derivative of medicinal polymer fpsdm and tp-5
Thing fixed point connects, and improves drug bioavailability, extends Half-life in vivo.Patented product of the present invention adopts structure single
Functional drug structural modification thing fpsdm, has effectively reached the effect to enzyme shielding, has fundamentally solved tp-5 half-life short
The shortcoming being only 30 seconds, effectively raises medicine long half time in vivo, enhances immunologic balance adjustment effect.The present invention's
Conjugates have important application prospect in New-type long-acting immunologic balance regulating drug research and development field.
Brief description
The vitro half-lives curve chart of the modified outcome Thymopentin methyl ester conjugates that Fig. 1 is prepared for the present invention and tp-5.
Specific embodiment
Below in conjunction with accompanying drawing, the principle of the present invention and feature are described, example is served only for explaining the present invention, and
Non- for limiting the scope of the present invention.
Embodiment 1
The chemosynthesis of intermediate z-fpsdm-arg (pbf)-lys (boc)-oh
Z-fpsdm-arg (pbf)-lys (boc)-ome is dissolved in meoh (18ml), stirring is lower to add lioh aqueous solution
(1.0m, 2.0ml).Reactant liquor is stirred at room temperature 3h to tlc display reaction completely.Pump solvent, residue in 30 DEG C of decompressions
It is dissolved in dcm (60ml), use hcl solution (0.2m, 2 × 20ml) and saturated aqueous common salt (2 × 20ml) washing afterwards successively, anhydrous
na2so4It is dried, after 30 DEG C of decompressions pump solvent, gained residue dcm/et2O/haxane (1:6:1) recrystallization, obtains
Target product z-fpsdm-arg (pbf)-lys (boc)-oh (mf.c66h108n10o20S, mw.1393.70).
Embodiment 2
The chemosynthesis of conjugates fpsdm-tp-5 derivant
By z-fpsdm-arg (pbf)-lys (boc)-oh 197.6mg (mf.c66h108n10o20S, mw.1393.70,
0.142mmol) it is dissolved in anhydrous dcm (10ml) with hoct 89.2mg (0.568mmol, 4eq), stirring is lower to add dic 26 μ l
(0.17mmol, 1.2eq), at room temperature after stirring 1h, is added drop-wise to tripeptide derivative h-asp (otbu)-val-tyr-ome
0.066g(c23h35n3o7, mw.465.55,0.142mmol) dcm (8ml) solution in, be simultaneously introduced diea 0.10ml, in room
Temperature is lower to stir 8h to tlc display reaction completely.Add dcm (50ml) in reactant liquor, use hcl (0.2m, 2 × 20ml), h successively2o
(20ml), naoh (0.2m, 2 × 20ml) and saturation nacl liquid (2 × 20ml) washing, use na afterwards2so4It is dried, reduce pressure in 30 DEG C
Pump solvent and obtain 0.188g conjugates crude product z-fpsdm-arg (pbf)-lys (boc)-asp (otbu)-val-tyr-ome
(mf.c89h141n13o26S, mw.1841.23), yield 76%.Add trifluoroacetic acid 5ml in crude product, and be stirred at room temperature
4h, with protection group boc-, otbu- of deprotection base z and functional side chain group and pbf-, pump three after 30 DEG C of decompressions
Fluoroethanoic acid, residue et2O (3 × 5ml) washs, and obtains conjugates fpsdm-arg-lys-asp-val-tyr-ome after being dried
(mf.c59h103n13o9, mw.1298.55) and 0.1327g, yield 72%, ms.1299.6 [(m+1)+].
Embodiment 3
Boc- protects the synthesis of dipeptidase derivant
By boc-val-oh (1-2, mw.217.27,1.0864g, 5.0mmol) and hoct (mf.c5h7n3o3,
Mw.157.12,0.9427g, 6.0mmol, 1.2eq) it is dissolved in anhydrous dcm (4.0ml), slowly Deca dic under stirring
(mf.c7h14n2, mw.126.20, d.0.839,0.91ml, 6.0mmol, 1.2eq) in above-mentioned solution.By h-tyr-ome
Hcl (1-1, mw.231.68,1.1584g, 5.0mmol) is suspended in dcm (5.0ml), and stirring is lower to add diea (mf.c8h19N,
Mw.129.24, d.0.728,1.95ml, 11.0mmol, 2.2eq), then by the mixed solution Deca of 1-2/hoct/dic/dcm
To wherein, stirring 2h under room temperature, tlc detection reaction is completely.Nahco is used after reacting liquid filtering3(0.2m, 2 × 25ml), hydrochloric acid
(0.2m, 2 × 20ml) and saturated aqueous common salt (30ml) extract successively, anhydrous na2so4It is dried, after concentrating under reduced pressure, obtain pale yellowish oil
Thing, plus et2O has urea to separate out, and fast filtering adds appropriate et2O places crystallization, obtains boc-val-tyr-ome
(mf.c19h30n2o5, mw.366.46, products nr: 1-3) and 1.4292g, product is white solid, yield 78%.Related data:
rf=0.76 (acoet/hexane=3/1).Mp:124-126.5 DEG C, hplc:3.583min (elution program: b-3),
ms.367.5[(m+1)+],384.4[(m+nh4)+], 389.5 [(m+na)+].
Add trifluoroacetic acid in described product (boc-val-tyr-ome), stir together, then carry out vacuum distillation and pump
Trifluoroacetic acid, obtains residue, then carries out described residue successively washing, is dried, that is, obtains the dipeptides h- of deprotection
Val-tyr-ome (mf, c15h22n2o4, mw.294.35, products nr: 1-5).Related data: rf=0.33 (acoet/
Hexane=2/1).
Embodiment 4
Fmoc protects the synthesis of tripeptide derivative
H-val-tyr-ome (1-5,0.2944g, 1.0mmol) and diea (0.22ml, 1.2mmol, 1.2eq) is dissolved in no
Water dcm (3.0ml).Fmoc-asp (otbu)-oh (0.4115g, 1.0mmol) and hoct (0.1885g, 1.2mmol, 1.2eq)
It is dissolved in dcm (8.0ml) slowly Deca dic (0.18ml, 1.2mmol, 1.2eq), after stirring 15min, drop to 1- with stirring
In 5 alkaline solution, react under room temperature to tlc monitoring reaction completely.Reactant liquor filters urea first, then uses nahco successively3Molten
Liquid (0.1m, 2 × 15ml), water (15ml), hydrochloric acid solution (0.2m, 2 × 15ml), saturation nacl solution (20ml) extraction, organic
Layer uses anhydrous na2so4It is dried, filter, after filtrate reduced in volume, plus et2O recrystallization obtains fmoc-asp (otbu)-val-tyr-
ome(mf.c38h45n3o9, mw.687.79, products nr: 1-6) and 0.6247g.Product is beige solid, yield 85%.Related
Data: rf=0.57 (acoet/hexane=1/1).Mp:141-143.5 DEG C, hplc:3.981min (elution program: b-3),
ms.688.7[(m+1)+], 710.7 [(m+na)+].
Fmoc-asp (otbu)-val-tyr-ome (1-6,0.3649g, 0.5mmol) is dissolved in diethylamine/dcm
(10%) (10ml) in solution, stir 6h under room temperature, show reaction completely to tlc monitoring.Reactant liquor pumps dcm in 30 DEG C of decompressions
And diethylamine, residue resulting solution after dcm dissolving is successively with water, nahco3(0.1m), water and saturated aqueous common salt extraction, no
Water na2so4After drying, drain and can get crude product.By crude product petroleum ether supersound washing, until fmoc group eliminates.Product
For grease h-asp (otbu)-val-tyr-ome (mf.c23h35n3o7, mw.465.55, products nr: 1-7) and 0.2002g, receives
Rate 86%.rf=0.43 (hexane/acoet=1/10).
Embodiment 5
Pbf protects the synthesis of dipeptidase derivant
fmoc-arg(pbf)-oh(c34h40n4o7S, mw.648.77,0.6488g, 1.0mmol) and diea (0.22ml,
1.2mmol, 1.2eq) it is dissolved in anhydrous dcm (5.0ml).h-lys(boc)-ome·hcl(mf.c12h24n2o4Hcl,
Mw.296.79,0.2968g, 1.0mmol) it is dissolved in dcm (3.0ml) and slowly drips with hoct (0.1885g, 1.2mmol, 1.2eq)
Plus dic (0.18ml, 1.2mmol, 1.2eq), drop in above-mentioned solution after stirring 15min, under room temperature, react anti-to tlc monitoring
Should be completely.Then use nahco successively3Solution (0.1m, 2 × 15ml), water (15ml), hydrochloric acid solution (0.2m, 2 × 15ml), full
With nacl solution (20ml) extraction, anhydrous na2so4It is dried, obtain fmoc-arg (pbf)-lys (boc)-ome (mf.c46h62n6o10,
Mw.891.09, products nr: 1-8) 0.7842g.Product is beige solid, yield 88%.Related data: rf=0.42
(acoet/hexane=2/1).Mp:136-138 DEG C, hplc:3.643min (elution program: b-3), ms.892.1 [(m+1
)+],909.1[(m+nh4)+], 914.1 [(m+na)+].
Embodiment 6
The synthesis of z-fpsdm-arg (pbf)-lys (boc)-ome
By the z-fpsdm 0.575g (mf.c in backbone and with carboxyl as functional group36h60n4o13；
Mw.756.89,0.76mmol) and hoct 0.238g (mf.c5h7n3o3, mw.157.12,1.52mmol, 2.0eq) be dissolved in anhydrous
Dcm (10ml), stirring is lower to add dic 0.137ml (0.895mmol, 1.2eq), forms activity at room temperature after stirring 45min
Ester b, is then added drop-wise to the dcm (6ml) of h--arg (pbf)-lys (boc)-ome 0.508g (mw.668.85,0.760mmol)
In solution, it is simultaneously introduced diea 0.20ml (mf.c8h19N, mw.129.24, d.0.728,1.12mmol), stir at room temperature
6h to tlc display reaction is completely.Add dcm (50ml) in reactant liquor, use hcl (0.2m, 2 × 20ml), h successively2o(20ml)、
Naoh (0.2m, 2 × 20ml) and saturation nacl solution (2 × 20ml) washing, use na afterwards2so4It is dried, pump in 30 DEG C of decompressions
Solvent obtains pale yellowish oil product z-fpsdm-arg (pbf)-lys (boc)-ome 0.887g (mf.c67h110n10o20S,
Mw.1407.73), yield 83%.Tlc display product is single speckle rfFor 0.53 (hexane:acoet:meoh=1:1:
0.5；uv254And i2Colour developing).
BIOLOGICAL ACTIVITY EXAMPLES
The embodiment 7 patent system of the present invention standby modified outcome stability study of resistance to enzymolysis in vitro
Experimental technique: prepare the tp-5 solution that initial protein concentration is 1.5mg/ml, the modification of preparation same molar ratio
Product fpsdm-tp-5 solution, prepare initial protein concentration be 1.5mg/ml trypsin solution, trypsin solution with treat
Survey polypeptide solution to mix with the ratio of 1:50, react 3min, 6min, 9min, 12min, 15min sampling respectively in 37 DEG C of water-baths and add
Excess of oxygen sodium hydroxide solution terminating reaction.Using rp-hplc gradient elution analysis, average in triplicate.
Experimental result: add trypsin digestion under the conditions of 37 DEG C of waters bath with thermostatic control, substantially, external enzymolysis partly declines for tp-5 degraded
Phase is (2.51 ± 0.36) min；And accurately modified outcome fpsdm-tp-5 then shows preferably resistance to enzymolysis stability, it is external
The enzymolysis half-life is that (18.76 ± 0.48) enzymolysis stability improves 7.47 times, and vitro half-lives curve chart is shown in Figure of description
1.
The activity research (zoopery) of the modified outcome of embodiment 8 present invention preparation
Experimental technique: from mice 60, body weight 19-21g, purchased from Shandong University's Experimental Animal Center.1. mice is pressed
Body weight is randomly divided into 3 groups, respectively blank group, tp-5 group, fpsdm-tp-5 group.Every group 10.2. the medication of tp-5 group is as follows: presses
The dosage lumbar injection of 0.2mg/ (kg d), successive administration 6 days；The 1st, 4 days lumbar injection 0.2mg/ (kg of fpsdm-tp-5 group
D) fpsdm-tp-5 normal saline solution；Blank group injects isopyknic normal saline daily.3. after last dose half an hour
Tail vein blood smear, with the dyeing of giemsa-weigh liquid, the cell number that microscopy T Lymphocyte Cell is activated, calculates and turns
Rate.4. carry out Culling heart blood after 1 hour in last dose and do Flos Rosae Rugosae experiment.5. put to death mice at cervical vertebra, aseptic taking-up is little
The thymus of Mus and spleen, calculate the thymus index of mice.
Table 1 Activity determination result
Experimental result: all of experimental data Statistics Application software spss 10.0 counts.As can be seen from Table 1
The periphery t lymhocyte transformation rate of fpsdm-tp-5 and tp-5, e- rosette forming rate and thymus index and blank group contrast
All there is significant difference (p < 0.05).Represent that the two all can improve the cellular immune function of mice；Fpsdm-tp-5 usage amount is more
Few, and effect is more preferable than tp-5, can play long-acting in vivo.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all spirit in the present invention and
Within principle, any modification, equivalent substitution and improvement made etc., should be included in protection scope of the present invention.
Claims (9)
1. a kind of accurate preparation method modifying Thymopentin methyl ester conjugates is it is characterised in that comprise the following steps:
1) z-fpsdm-arg (pbf)-lys (boc)-oh and 1- hydroxyl -1h-1,2,3- triazole -4- carboxylic acid, ethyl ester are dissolved in anhydrous
In dichloromethane, add n while stirring, n- DIC is reacted, and obtains active ester a；
2) described active ester a is added drop-wise in the alkaline solution of tripeptide derivative, is simultaneously introduced n- diisopropylethylamine, stirs together
Mix to reacting completely, then add dichloromethane, and carry out successively washing, be dried, vacuum distillation, obtain crude product thymus five
Peptide methyl ester conjugates；
3) add trifluoroacetic acid in described crude product, stir together, then carry out vacuum distillation and pump trifluoroacetic acid, remained
Then described residue is carried out washing, is dried, that is, obtain described Thymopentin methyl ester conjugates by thing successively.
2. preparation method according to claim 1 is it is characterised in that the time of stirring described in step 1) is 45～75
Minute.
3. preparation method according to claim 1 is it is characterised in that in step 2) in, described alkaline solution is dichloromethane
Alkane solution；The time of described stirring is 7～9 hours；Described washing step is with hcl solution, aqueous solution, naoh solution and saturation
Nacl solution is washed successively；Described desiccant is na2so4Desiccant；Temperature under described decompression is 20～35 DEG C.
4. preparation method according to claim 1 is it is characterised in that in step 3), the time of described stirring is 3～5
Hour；Described washing cleaning mixture used is et2o；Temperature under described decompression is 20～35 DEG C.
5. the preparation method according to any one of Claims 1-4 it is characterised in that described z-fpsdm-arg (pbf)-
Lys (boc)-oh is prepared via a method which:
1) by z-fpsdm and 1- hydroxyl -1h-1,2,3- triazole -4- carboxylic acid, ethyl esters are dissolved in anhydrous methylene chloride, while stirring plus
Enter n, n- DIC is reacted, and obtains active ester b；
2) described active ester b is added drop-wise in the alkaline solution of h-arg (pbf)-lys (boc)-ome, is simultaneously introduced n- diisopropyl
Base ethamine, is stirred together to and reacts completely；Add dichloromethane, then carry out successively again washing, be dried, vacuum distillation, obtain
To z-fpsdm-arg (pbf)-lys (boc)-ome；
3) above-mentioned gained z-fpsdm-arg (pbf)-lys (boc)-ome is added in meoh, adds lioh water while stirring
Solution is to reacting completely, then carries out vacuum distillation, pumps solvent, obtains residue；Described residue carried out successively wash, do
After dry, vacuum distillation, recrystallization, that is, synthesize described z-fpsdm-arg (pbf)-lys (boc)-oh.
6. preparation method according to claim 5 is it is characterised in that in step 1), and time of described stirring is 30～
60 minutes.
7. preparation method according to claim 5 is it is characterised in that in step 2) in, described alkaline solution is dichloromethane
Alkane solution；The time of described stirring is 5～7 hours；The step of described washing is with hcl solution, aqueous solution, naoh solution and to satisfy
Washed successively with nacl solution；Desiccant used by described drying is na2so4Desiccant；Temperature under described decompression is 20
～35 DEG C.
8. preparation method according to claim 5 is it is characterised in that in step 3), the time of described stirring is 2～4
Hour；Described reduce pressure twice under temperature be 20～35 DEG C；The step of described washing be washed with hcl solution after, then with full
Washed with saline solution；Desiccant used by described drying is na2so4Desiccant；The dichloro that the material of described recrystallization is
Methane, et2O and normal hexane, wherein said dichloromethane, et2The ratio of weight and number of o and normal hexane is 1:6:1.
9. the Thymopentin methyl ester conjugates that the preparation method described in a kind of any one according to claim 1 to 8 is obtained.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610806260.7A CN106366177A (en) | 2016-09-07 | 2016-09-07 | Thymopentin methyl ester conjugate and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610806260.7A CN106366177A (en) | 2016-09-07 | 2016-09-07 | Thymopentin methyl ester conjugate and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106366177A true CN106366177A (en) | 2017-02-01 |
Family
ID=57898951
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610806260.7A Pending CN106366177A (en) | 2016-09-07 | 2016-09-07 | Thymopentin methyl ester conjugate and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106366177A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102558301A (en) * | 2011-12-13 | 2012-07-11 | 烟台海安多肽药物研发技术有限公司 | Functional precision structure drug modifier-thymopetidum acetyl methyl ester conjugate (FPSDM-TP5) and preparation method thereof |
| CN103103239A (en) * | 2013-01-22 | 2013-05-15 | 华南农业大学 | Method for synthesizing thymopentin by enzyme |
| CN103242431A (en) * | 2013-05-20 | 2013-08-14 | 齐鲁制药有限公司 | Preparation method of bivalirudin |
| CN103450346A (en) * | 2012-09-14 | 2013-12-18 | 烟台海安多肽药物研发技术有限公司 | Liquid-phase synthesis method of eptifibatide |
| CN104672301A (en) * | 2014-12-11 | 2015-06-03 | 烟台海安药物研发有限公司 | Thymopetidum acetyl methyl ester conjugate (FPSDM-TP5) and preparation method thereof |
-
2016
- 2016-09-07 CN CN201610806260.7A patent/CN106366177A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102558301A (en) * | 2011-12-13 | 2012-07-11 | 烟台海安多肽药物研发技术有限公司 | Functional precision structure drug modifier-thymopetidum acetyl methyl ester conjugate (FPSDM-TP5) and preparation method thereof |
| CN103450346A (en) * | 2012-09-14 | 2013-12-18 | 烟台海安多肽药物研发技术有限公司 | Liquid-phase synthesis method of eptifibatide |
| CN103103239A (en) * | 2013-01-22 | 2013-05-15 | 华南农业大学 | Method for synthesizing thymopentin by enzyme |
| CN103242431A (en) * | 2013-05-20 | 2013-08-14 | 齐鲁制药有限公司 | Preparation method of bivalirudin |
| CN104672301A (en) * | 2014-12-11 | 2015-06-03 | 烟台海安药物研发有限公司 | Thymopetidum acetyl methyl ester conjugate (FPSDM-TP5) and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 姜凤超 编著: "《药物合成》", 30 September 2008, 化学工业出版社 * |
| 朱彬 编著: "《有机合成》", 31 January 2014, 西南交通大学出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100427502C (en) | Antineoplastic oligopeptide and its preparation method and application | |
| CN101541743A (en) | Transdermal delivery systems of peptides and related compounds | |
| CN104288779B (en) | PEGization peptides dendrimer targeting drug delivery system of load gemcitabine and preparation method thereof | |
| CN104370862B (en) | Water-soluble antitumor compound | |
| JPH06501950A (en) | Cyclic peptides, their preparation and their use as pharmacological compositions | |
| CN103204901A (en) | Preparation method for bifunctional conjugate | |
| CN104530199A (en) | Antitumor polypeptide, and preparation method and application thereof | |
| CN113929761B (en) | Novel somatostatin analogue peptide reconstruction and dimerization preparation and application thereof | |
| CN101870732B (en) | Method for synthesizing mono pegylation-thymopentin by solid phase and liquid phase combination | |
| CN102558298A (en) | Method for synthesizing tetrapeptide isomers by using solid phase peptide synthesis method and applications of tetrapeptide isomers | |
| CN106631957A (en) | Antitumor compound targeting FAP-alpha enzyme and preparation method and application thereof | |
| CN109908363B (en) | Targeted traceless release drug conjugate and preparation method and application thereof | |
| CN111068068A (en) | RGD polypeptide-camptothecin polypeptide drug conjugate and application thereof | |
| CN106366177A (en) | Thymopentin methyl ester conjugate and preparation method thereof | |
| CN102040569A (en) | Carotinoid derivatives and preparation method and application thereof | |
| US7183430B2 (en) | Compounds which can block the response to chemical substances or thermal stimuli or mediators of inflammation of nociceptors, production method thereof and compositions containing same | |
| CN107854693A (en) | The anticancer conjugate of integrin receptor target | |
| CN107298708A (en) | A kind of glucagon-like-peptide-1 with ehter bond（GLP-1）Analog and its application | |
| CN103204893A (en) | Method of improving oral absorption of carboxylic acids drugs by using Pep T1 targeting mimic-peptides diester prodrug technique | |
| CN106188273A (en) | Paclitaxel vapreotide conjugates and preparation method thereof | |
| CN102558308B (en) | Carrier polypeptide for forming medical compositions and preparation method and application of carrier polypeptide | |
| JPS61130236A (en) | Medicine containing fully synthetic alpha-peptide promoting exhaustion of sodium from human pericardium | |
| CN109721653A (en) | A kind of glucagon-like-peptide-1 fragment analogue and its application | |
| CN112094321A (en) | His-Gly-Glu modified methotrexate, synthesis, anti-transfer activity and application thereof | |
| CN103665106A (en) | Antineoplastic polypeptide and preparation method and applications thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 264006, Yantai, Shandong, Yantai Shandong Development Zone, Zhujianglu Road 32, Yantai Overseas Students Pioneer Park, No. 3 workshop, E District, room 539 Applicant after: Shandong Bio Technology Co., Ltd. Address before: 264006, Yantai, Shandong, Yantai Shandong Development Zone, Zhujianglu Road 32, Yantai Overseas Students Pioneer Park, No. 3 workshop, E District, room 539 Applicant before: Hai'an, Yantai medicament research and development company limited |
|
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170201 |