CN116496347A - Method for solid phase synthesis of pentapeptide - Google Patents

Method for solid phase synthesis of pentapeptide Download PDF

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CN116496347A
CN116496347A CN202310063155.9A CN202310063155A CN116496347A CN 116496347 A CN116496347 A CN 116496347A CN 202310063155 A CN202310063155 A CN 202310063155A CN 116496347 A CN116496347 A CN 116496347A
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compound
formula
reaction
dmf
butoxycarbonyl
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高俊
齐艳艳
林凡城
水祥飞
陈昊
孔祥林
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Jiangsu Hengrui Medicine Co Ltd
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    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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Abstract

The present disclosure relates to a method of solid phase synthesis of pentapeptides. Specifically, a solid phase synthesis method of 4-amino-N- [ N ] is provided 2 - [ N- [ N- [ N- ((R) -2-phenylpropyl) glycyl ]]-D-phenylalanyl]-D-leucyl]-D-lysyl]A process for the preparation of piperidine-4-carboxylic acid or a pharmaceutically acceptable salt thereof, which comprises the step of reacting a compound of formula I-8 with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid to form a compound of formula I-10, which process is high in yield and excellent in sample quality and is suitable for large scale production.

Description

Method for solid phase synthesis of pentapeptide
Technical Field
The present disclosure relates to a method of solid phase synthesis of pentapeptides.
Background
4-amino-N- [ N 2 - [ N- [ N- [ N- ((R) -2-phenylpropyl) glycyl ]]-D-phenylalanyl]-D-leucyl]-D-lysyl]Piperidine-4-carboxylic acid or pharmaceutically acceptable salts thereof are a class of Kappa Opioid Receptor (KOR) agonists of polypeptides. Clinical studies of both pain and itch have been conducted, with the pain indication having entered clinical stage III.
WO2017211272 reports a process for the synthesis of the aforementioned polypeptides using the "pooling" method, which essentially comprises the following steps,
wherein, the compound 1m and the compound 1f are prepared by multi-step reaction, the whole process route is longer, the yield is lower, the product quality is difficult to ensure, and the method is not suitable for large-scale production. Therefore, there is an urgent need to search for a new process route suitable for mass production needs.
Disclosure of Invention
The present disclosure provides a process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof,
the process comprises the step of reacting a compound of formula I-8 with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid to form a compound of formula I-10,
wherein P is 1 、P 2 And P 3 Each is an amino protecting group.
In some embodiments, P 1 、P 2 And P 3 Each is tert-butoxycarbonyl (Boc).
In some embodiments, a method of preparing a compound of formula I or a pharmaceutically acceptable salt thereof comprises the step of reacting a compound of formula I-8a with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid to form a compound of formula I-10a,
in other embodiments, the process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof comprises the steps of removing Fmoc (9-fluorenylmethoxycarbonyl) from a compound of formula I-8 under piperidine conditions, followed by condensation with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid or a salt thereof to form a compound of formula I-10, the condensation reagent being selected from the group consisting of benzotriazole-N, N, N ', N' -tetramethylurea Hexafluorophosphate (HBTU), 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylethylamine (DIEA),
in some embodiments, the molar ratio of benzotriazol-N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HBTU), 1-Hydroxybenzotriazole (HOBT), and N, N-Diisopropylethylamine (DIEA) is 0.8-1.2:0.8-1.2.
In some embodiments, the molar ratio of benzotriazole-N, N' -tetramethyluronium Hexafluorophosphate (HBTU), 1-Hydroxybenzotriazole (HOBT), and N, N-Diisopropylethylamine (DIEA) is 1:1:1.
In some embodiments, the molar amount of benzotriazol-N, N' -tetramethyluronium Hexafluorophosphate (HBTU) is 2.0 to 3.0 times that of the compound of formula I-8, including values between 2.0, 2.2, 2.4, 2.6, 2.8, 3.0 times or any two.
In some embodiments, the condensation reaction temperature is 15 to 45 ℃, including 15 ℃, 17 ℃, 19 ℃, 21 ℃, 23 ℃, 25 ℃, 27 ℃, 29 ℃, 31 ℃, 33 ℃, 35 ℃, 37 ℃, 39 ℃, 41 ℃, 43 ℃, 45 ℃, or values between any two.
Further, in other embodiments, the compound of formula I-8 is reacted with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid in a "one-pot" process to form the compound of formula I-10, or the conversion of the compound of formula I-8 to the compound of formula I may be a one-step or multi-step reaction, which may be a "one-pot multi-step" or "one-pot" process if a multi-step reaction.
In another aspect, the process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof further comprises the step of reacting a compound of formula I-10 under acidic conditions to form a compound of formula I,
wherein P is 1 And P 2 Is an amino protecting group.
In some embodiments, the acidic reagent is a 50-100% trifluoroacetic acid solution. In other embodiments, there is also triisopropylsilane in the trifluoroacetic acid solution. In some embodiments, the acidic reagent is selected from 95% trifluoroacetic acid/2.5% triisopropylsilane/2.5% water (v/v/v). The triisopropylsilane is added and utilized to obtain a cleaner product, which is beneficial to subsequent purification.
In some embodiments, the aforementioned cleavage reaction temperature is 15-30 ℃, including 15 ℃, 17 ℃, 19 ℃, 21 ℃, 23 ℃, 25 ℃, 27 ℃, 29 ℃, 30 ℃, or values between any two.
Some embodiments provide a process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof comprising the steps of removing Fmoc of formula I-8a under piperidine conditions, followed by condensation with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid or a salt thereof under 1-hydroxybenzotriazole and N, N' -diisopropylcarbodiimide conditions to form a compound of formula I-10a, and reacting the compound of formula I-10a under acidic conditions to form a compound of formula I,
in another aspect, the present disclosure provides a process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof, further comprising the step of removing Fmoc from a compound of formula I-6 under piperidine (PIP) conditions, followed by condensation with Fmoc-D-Phe-OH (a compound of formula I-7) to form a compound of formula I-8, the condensation reagent being selected from the group consisting of 1-hydroxybenzotriazole and N, N' -diisopropylcarbodiimide,
in some embodiments, the molar ratio of 1-hydroxybenzotriazole to N, N' -diisopropylcarbodiimide is from 0.8 to 1.2:0.8 to 1.2.
In some embodiments, the molar ratio of 1-hydroxybenzotriazole to N, N' -diisopropylcarbodiimide is 1:1.
In some embodiments, the aforementioned condensation reaction temperature is 15 to 45 ℃, including 15 ℃, 17 ℃, 19 ℃, 21 ℃, 23 ℃, 25 ℃, 27 ℃, 29 ℃, 31 ℃, 33 ℃, 35 ℃, 37 ℃, 39 ℃, 41 ℃, 43 ℃, 45 ℃, or values between any two.
In some embodiments, the conversion of the compound of formula I-6 to the compound of formula I-8 may be a one-step or multi-step reaction, which if a multi-step reaction may be a "one-pot multi-step process" or a "one-pot process".
In another aspect, the compound of formula I-6 is prepared by a process comprising the steps of reacting a resin (2-chlorotrityl chloride resin, 2-CTC) with a compound of formula I-1, converting the compound of formula I-2 to a compound of formula I-4 by reaction with Fmoc-D-Lys (Boc) -OH, and converting the compound of formula I-4 to a compound of formula I-6 by reaction with Fmoc-D-Leu-OH,
(Fmoc-D-Lys (Boc) -OH, formula I-3 a),(Fmoc-D-Lys (Boc) -OH, formula I-3 a), -A>(Fmoc-D-Leu-OH，I-5)。
In other embodiments, the process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof further comprises the step of purifying the crude compound of formula I by a C8 packed chromatography column, wherein the mobile phase is 0.1% ammonia/acetonitrile, gradient elution.
Some embodiments provide a process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof, comprising the steps of,
step 1) under the condition of N, N-diisopropylethylamine, reacting a compound I-1 with a 2-CTC resin to form a compound I-2;
step 2) Fmoc removal of compound I-2 under 20% PIP/DMF followed by reaction with compound I-3a under DIC/HOBT conditions to form compound I-4a;
step 3) Fmoc removal of compound I-4a under 20% PIP/DMF followed by reaction with compound I-5 under DIC/HOBT conditions to form compound I-6a;
step 4) Fmoc removal of compound I-6a under 20% PIP/DMF followed by reaction with compound I-7 under HBTU/HOBT/DIEA conditions to form compound I-8a;
step 5) Fmoc removal of compound I-8a under 20% PIP/DMF followed by reaction with compound I-9a under DIC/HOBT conditions to form compound I-10a;
step 6) Compound I-10a in TFA/TIS/H 2 O (V: v=95:2.5:2.5) to form the target compound.
The present disclosure also provides compounds of formula I-10,
wherein P is 1 And P 2 Each is an amino protecting group. In some embodiments, P 1 And P 2 Each is tert-butoxycarbonyl (Boc).
In some embodiments, the compounds of formula I-10 are
The present disclosure also provides a pharmaceutical composition comprising an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof prepared by the foregoing process and one or more pharmaceutically acceptable excipients.
In another aspect, the present disclosure also provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, as described above, in the manufacture of a medicament for the treatment of a condition associated with a KOR receptor selected from the group consisting of pain, inflammation, itch, edema, hyponatremia, hypokalemia, ileus, cough, and glaucoma, selected from the group consisting of neuropathic pain, somatic pain, visceral pain, skin pain, arthritic pain, kidney stone pain, uterine cramps, dysmenorrhea, endometriosis, dyspepsia, post-surgical pain, post-medical treatment pain, ocular pain, otitis pain, breakthrough cancer pain, and pain associated with GI disorders.
In another aspect, the present disclosure also provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, as described above, in the manufacture of a medicament for the treatment of a pain selected from neuropathic pain, somatic pain, visceral pain, skin pain, arthritic pain, kidney stone pain, uterine cramps, dysmenorrhea, endometriosis, dyspepsia, post-surgical pain, post-medical treatment pain, ocular pain, otitis pain, breakthrough cancer pain, and pain associated with GI disorders.
The methods of the present disclosure also include post-treatment operations such as one or more of filtration, extraction, concentration, column chromatography to obtain the pure target product.
The pharmaceutically acceptable salts of the present disclosure are the product of the compound of formula I with an acid selected from, but not limited to, hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, malic acid, oxalic acid, and the like.
The salts of the present disclosure are the products of the corresponding compounds salified with an acid selected from, but not limited to, hydrochloric acid, methanesulfonic acid, trifluoromethanesulfonic acid, phosphoric acid, oxalic acid, and the like.
The terms used in the present disclosure have the following meanings, unless stated to the contrary:
the present disclosure "to form" and "convert to" does not specifically refer to the conversion reaction between two substrates being a single step, and may be a single step or a multi-step reaction between two substrates. If the intermediate contains an amino protecting group, the intermediate is subjected to a one-step deamination protecting agent and then reacted with a corresponding substrate to obtain a corresponding target product.
"alkyl" refers to a saturated aliphatic hydrocarbon group comprising from 1 to 6 carbon atoms. Non-limiting examples include, but are not limited to, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclopentyl, and the like.
"amino protecting groups" are suitable groups for amino protection known in the art, see literature ("Protective Groups in Organic Synthesis", 5) Th Ed.T.W.Greene&P.g.m.wuts). Examples include, but are not limited to, urethane protecting groups such as t-butoxycarbonyl (Boc), allyloxycarbonyl (Alloc), 9-fluorenylmethoxycarbonyl (Fmoc), and benzyloxycarbonyl (Cbz).
The term "therapeutically effective amount" or "effective amount" as used in this disclosure includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount is also meant to be an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on the following factors: such as the condition to be treated, the general health of the patient, the route of administration and the dosage and severity of the side effects. An effective amount may be the maximum dose or regimen that avoids significant side effects or toxic effects.
The "excipient" as described in this disclosure includes, but is not limited to, any auxiliary agent, carrier, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizing agent, isotonic agent, or emulsifying agent that has been approved by the U.S. food and drug administration for use in humans or livestock animals.
The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift (. Delta.) of 10 -6 Units of (ppm) are given. NMR was performed using Bruker AVANCE-400 nuclear magnetic resonance apparatus with deuterated dimethyl sulfoxide (DMSO-d) 6 ) Deuterated chloroform (CDCl) 3 ) Deuterated methanol (CD) 3 OD), internal standard is Tetramethylsilane (TMS).
HPLC detection: chromatography columns packed with octadecylsilane chemically bonded silica (e.g. Phenomenex Gemini C18)150×4.6mm,5 μm), phosphate buffer/acetonitrile as mobile phase, gradient elution.
The preparation liquid phase can adopt CS-Prep200 preparation chromatographic system, chromatographic column with C8 as filler, 0.1% ammonia water/acetonitrile as mobile phase, and gradient elution.
Detailed Description
The present disclosure will be described in detail below in conjunction with specific embodiments, which are only for illustrating the technical solutions of the present disclosure and not limiting the present disclosure in any way, so that those skilled in the art will more fully understand the present disclosure.
Example 1
Step 1:
10g of 2-CTC resin (substitution degree: 0.98 mmol/g) was weighed, charged into a reactor, DMF (100 mL) was added, N 2 Bubbling reaction for 0.5-1h, filtering and draining.
Weighing compound 1 (5.6 g), adding DMF (48 mL) and DCM (72 mL), dissolving, adding DIEA (6.5 g), stirring, mixing, adding into a reactor, and adding N 2 Bubbling reaction is carried out for 18-22 h, filtering, washing and drying are carried out, thus obtaining the compound 2.
The yield calculation formula:
the yield thereof was found to be 93.7%.
Step 2:
weigh Compound 2 (15 g) and add to the reactor, DMF (100 mL), N 2 Bubbling reaction for 0.5-1h, filtering and draining.
20% piperidine/DMF (100 mL) was added to the reaction vessel, stirred at 15-40℃until the reaction was complete, filtered, DMF washed and filtered to dryness.
Compound 3 (11 g), HOBT (3.2 g), HBTU (9.0 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (6.1 g) were added, stirred and dissolved, then added to a reactor, the reaction was stirred and completely filtered under nitrogen bubbling at 15-40℃and the DMF was washed and filtered to dryness to give Compound 4.
Step 3:
20% piperidine/DMF (100 mL) was added to the previous reactor and the reaction was stirred to completion with nitrogen sparge at 15-40 ℃, filtered, DMF washed and filtered to dryness.
Compound 5 (8.3 g), HOBT (3.2 g), HBTU (9.0 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (6.1 g) were added, stirred and dissolved, then added to a reactor, reacted to completion under stirring with nitrogen bubbling at 15-40 ℃, filtered, washed and filtered to dryness to give compound 6.
Step 4:
20% piperidine/DMF (100 mL) was added to the reactor and the reaction was stirred at 15-40℃until complete, filtered, DMF washed and filtered to dryness.
Compound 7 (9.1 g), HOBT (3.2 g), HBTU (9.0 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (6.1 g) were added, stirred and dissolved, then added to a reactor, stirred and reacted at 15-40 ℃ until completion, filtered, washed and filtered to dryness to give compound 8.
Step 5:
20% piperidine/DMF (100 mL) was added to the reactor and the reaction was stirred at 15-40℃until complete, filtered, washed and filtered to dryness.
Compound 9 (6.9 g), HOBT (3.2 g) and HBTU (9.0 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (6.1 g) were added, stirred and dissolved, then added into a reactor, stirred and reacted at 15-40 ℃ until complete, filtered, washed, filtered until dry, and dried in a vacuum oven for 12-18 h to obtain compound 10.
Step 6:
TFA/TIS/H 2 The O (V: V: V=95:2.5:2.5) solution is added into a reaction kettle, and the dried compound 10 is added, and the reaction is stirred at 15-30 ℃ until the completion. The reaction mixture was filtered, slowly poured into isopropyl ether, filtered and washed. And (3) filtering to dryness, and drying the obtained solid in a vacuum drying oven for 12-18 hours to obtain the target compound with the HPLC purity of 87.7%.
10g of 2-CTC resin (substitution degree: 0.98 mmol/g) was weighed, charged into a reactor, DMF (100 mL) was added, N 2 Bubbling reaction for 0.5-1h, filtering and draining.
Weighing compound 1 (5.6 g), adding DMF (48 mL) and DCM (72 mL), dissolving, adding DIEA (6.5 g), stirring, mixing, adding into a reactor, and adding N 2 Bubbling reaction is carried out for 18-22 h, filtering, washing and drying are carried out, thus obtaining the compound 2.
The yield calculation formula:
the yield thereof was found to be 94.1%.
Step 2:
weigh Compound 2 (15 g) and add to the reactor, DMF (100 mL), N 2 Bubbling reaction for 0.5-1h, filtering and draining.
20% piperidine/DMF (100 mL) was added to the reaction vessel, stirred at 15-40℃until the reaction was complete, filtered, DMF washed and filtered to dryness.
Compound 3 (11 g), DEPBT (8.9 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (7.8 g) were added, stirred and dissolved, then added to a reactor, the reaction was stirred under nitrogen bubbling at 15-40 ℃ and filtered completely, and the mixture was washed with DMF and filtered to dryness to give Compound 4.
Step 3:
20% piperidine/DMF (100 mL) was added to the previous reactor and the reaction was stirred to completion with nitrogen sparge at 15-40 ℃, filtered, DMF washed and filtered to dryness.
Compound 5 (8.3 g), debt (8.9 g), DMF (50 mL), DCM (50 mL) and DIEA (7.8 g) were weighed, added to the reactor after stirring to dissolve, reacted to completion with stirring under nitrogen bubbling at 15-40 ℃, filtered, washed, and filtered to dryness to give compound 6.
Step 4:
20% piperidine/DMF (100 mL) was added to the reactor and the reaction was stirred at 15-40℃until complete, filtered, DMF washed and filtered to dryness.
Compound 7 (9.1 g), debt (8.9 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (7.8 g) were added, stirred and dissolved, then added to a reactor, stirred and reacted at 15-40 ℃ until completed, filtered, washed and filtered to dryness to give compound 8.
Step 5:
20% piperidine/DMF (100 mL) was added to the reactor and the reaction was stirred at 15-40℃until complete, filtered, washed and filtered to dryness.
Compound 9 (6.9 g) and debt (8.9 g) were weighed, DMF (50 mL), DCM (50 mL) and DIEA (7.8 g) were added, stirred and dissolved, then added into a reactor, stirred and reacted at 15-40 ℃ until complete, filtered, washed, filtered until dry, and dried in a vacuum oven for 12-18 h to give compound 10.
Step 6:
TFA/TIS/H 2 The O (V: V: V=95:2.5:2.5) solution is added into a reaction kettle, and the dried compound 10 is added, and the reaction is stirred at 15-30 ℃ until the completion. The reaction mixture was filtered, slowly poured into isopropyl ether, filtered and washed. And (3) filtering to dryness, and drying the obtained solid in a vacuum drying oven for 12-18 hours to obtain the target compound with the HPLC purity of 88.5%.
Example 3
Step 1:
1000g of 2-CTC resin (substitution degree 0.5-1.0 mmol/g) was weighed, put into a glass reaction vessel, DMF (12L), N was added 2 Bubbling reaction for 0.5-1h, filtering and draining.
Weighing compound 1 (560 g), adding DMF (4.8L) and DCM (7.2L), dissolving, adding DIEA (648 g), stirring, mixing, adding into a reactor, and adding N 2 Bubbling reaction is carried out for 18-22 h, filtering, washing and drying are carried out, thus obtaining the compound 2.
The yield calculation formula:
the yield thereof was found to be 94.5%.
Step 2:
weighing compound 2 (1500 g) and adding into a glass reaction kettle, adding DMF (12L) and N 2 Bubbling reactionFiltering and draining for 0.5-1 h.
20% piperidine/DMF (12L) was added to the reaction vessel and stirred at 15-40℃until the reaction was complete, filtered, DMF was washed and filtered to dryness.
Weighing compound 3 (1100 g) and HOBT (320 g), adding DMF (6L), DCM (6L) and DIC (298 g), stirring for dissolution, adding into a glass reaction kettle, stirring for complete reaction under 15-40 ℃ reaction nitrogen bubbling, filtering, washing with DMF, and filtering to obtain compound 4.
Step 3:
20% piperidine/DMF (12L) was added to the previous glass reactor and the reaction was stirred at 15-40℃with nitrogen bubbling until complete, filtered, DMF washed and filtered to dryness.
Weighing compound 5 (833 g), HOBT (320 g) in a configuration barrel, respectively adding DMF (6L), DCM (6L) and DIC (298 g), stirring for dissolving, adding into a glass reaction kettle, stirring for reacting at 15-40 ℃ under nitrogen bubbling until the reaction is complete, filtering, washing, and filtering to obtain the compound 6.
Step 4:
20% piperidine/DMF (12L) was added to a glass reactor and reacted to completion with stirring at 15-40℃and filtered, DMF was washed and filtered to dryness.
Weighing compound 7 (915 g) and HOBT (320 g) in a configuration barrel, respectively adding DMF (6L), DCM (6L) and DIC (298 g), stirring for dissolving, adding into a glass reaction kettle, stirring for reaction at 15-40 ℃ until the reaction is completed, filtering, washing, and filtering to obtain the compound 8.
Step 5:
20% piperidine/DMF (12L) was added to a glass reactor and reacted to completion with stirring at 15-40℃and filtered, washed and filtered to dryness.
Weighing a compound 9 (690 g), HOBT (320 g) and HBTU (895 g) in a configuration barrel, adding DMF (6L), DCM (6L) and DIEA (610 g), stirring for dissolving, adding into a glass reaction kettle, stirring for reacting at 15-40 ℃ until the reaction is complete, filtering, washing, filtering to dryness, and drying in a vacuum drying oven for 12-18 h to obtain the compound 10.
Step 6:
TFA/TIS/H 2 O(V:V:V＝95:2.5:2.5 Adding the solution into a reaction kettle, stirring uniformly, and cooling to 0-15 ℃. Adding the dried compound 10, stirring at 15-30 ℃ to complete the reaction. Filtering, slowly pouring the reaction solution into isopropyl ether, filtering, washing, filtering to dryness, and drying the obtained solid in a vacuum drying oven for 12-18 hours to obtain 830g of target compound with the yield of 69% and the HPLC purity of 92.0%.
Dissolving the product obtained in the previous step in 0.2mol/L ammonium acetate solution to prepare a solution with the concentration of about 10mg/ml, filtering, preparing liquid phase by adopting a CS-Prep200 preparation chromatographic system and a chromatographic column with C8 as a filler, concentrating and freeze-drying to obtain 439g of a pure product, wherein the total yield is 52%, and the HPLC purity is 99.42%.

Claims (10)

1. A process for preparing a compound of formula I or a pharmaceutically acceptable salt thereof,
comprising the step of reacting a compound of formula I-8 with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid to form a compound of formula I-10,
wherein P is 1 、P 2 And P 3 Each is an amino protecting group, preferably t-butoxycarbonyl (Boc).
2. The process of claim 1, comprising the step of removing Fmoc from the compound of formula I-8 under piperidine conditions, followed by condensation with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid or a salt thereof to form the compound of formula I-10, the condensation reagent being selected from the group consisting of benzotriazole-N, N' -tetramethylurea hexafluorophosphate, 1-hydroxybenzotriazole and N, N-diisopropylethylamine.
3. A process according to claim 1 or 2, wherein the compound of formula I-8 is reacted with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid in a "one pot" process to form the compound of formula I-10.
4. The method of any one of claim 1 to 3, further comprising the step of reacting the compound of formula I-10 under acidic conditions to form the compound of formula I,
wherein P is 1 、P 2 And P 3 Is an amino protecting group.
5. The process according to claim 4, wherein the acidic reagent is 50-100% trifluoroacetic acid solution, preferably said trifluoroacetic acid solution is further provided with triisopropylsilane; further, the acidic reagent is preferably 95% trifluoroacetic acid/2.5% triisopropylsilane/2.5% water (v/v/v).
6. A process according to any one of claims 1 to 5, comprising the steps of removing Fmoc of formula I-8a under piperidine conditions, followed by condensation with (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid or a salt thereof under 1-hydroxybenzotriazole and N, N' -diisopropylcarbodiimide conditions to form a compound of formula I-10a, and reacting the compound of formula I-10a under acidic conditions to form a compound of formula I,
7. the process according to any one of claims 1-5, further comprising the step of removing Fmoc from the compound of formula I-6 under piperidine conditions, followed by condensation with Fmoc-D-Phe-OH (compound of formula I-7) to form the compound of formula I-8, the condensation reagent being selected from the group consisting of 1-hydroxybenzotriazole and N, N' -diisopropylcarbodiimide,
8. the process of any one of claims 1-7, further comprising the step of purifying the crude compound of formula I via a C8 packed chromatography column, wherein the mobile phase is 0.1% aqueous ammonia/acetonitrile.
9. The compound of the formula I-10,
wherein P is 1 、P 2 And P 3 Each is an amino protecting group, preferably t-butoxycarbonyl (Boc).
10. A pharmaceutical composition comprising an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof, prepared by the process of any one of claims 1-8, and one or more pharmaceutically acceptable excipients.
CN202310063155.9A 2022-01-25 2023-01-19 Method for solid phase synthesis of pentapeptide Pending CN116496347A (en)

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