CN116925105A

CN116925105A - Solid salt forms, crystalline forms, and methods of making, compositions, and uses of opioid receptor antagonist conjugates

Info

Publication number: CN116925105A
Application number: CN202211262369.0A
Authority: CN
Inventors: 何玫
Original assignee: Shanghai Hanmai Biopharmaceutical Technology Co ltd
Current assignee: Shanghai Hanmai Biopharmaceutical Technology Co ltd
Priority date: 2022-04-11
Filing date: 2022-10-14
Publication date: 2023-10-24
Also published as: WO2023197934A1

Abstract

The invention belongs to the technical field of medicine crystals, and relates to solid salt type and crystal type of an opioid receptor antagonist conjugate, and a preparation method, a composition and application thereof. Specifically, the conjugate has a structure shown in a formula I, and can form a solid phosphate which exists stably, wherein the molar ratio of phosphoric acid to the compound in the formula I is 1:1-3:1. The phosphate can be amorphous or have polymorphs of crystal forms 1, 2, 3 or 4, and the preparation method is simple and easy to implement, and is suitable for preventing and treating intestinal dysfunction caused by opioid, such as constipation.

Description

Solid salt forms, crystalline forms, and methods of making, compositions, and uses of opioid receptor antagonist conjugates

Technical Field

The invention belongs to the technical field of medicine crystals, and particularly relates to a solid salt type and a crystal type of an opioid receptor antagonist conjugate (especially a double opioid receptor antagonist conjugate), a preparation method, a medicine composition and medical application thereof.

Background

Cancer pain is one of the most common complications of progression of cancer to middle and late stages, with pain incidence of up to 75% or more in patients with advanced cancer. Cancer pain is a significant hazard to cancer patients, families and society.

Opium is the oldest analgesic, is the most effective analgesic so far, and has the advantages of strong analgesic effect, no organ toxicity effect after long-term administration, and the like. When pain in patients is aggravated by tumor progression, the analgesic effect can be enhanced by increasing the drug dose, so opioid analgesics have an irreplaceable position for patients with moderate and severe cancer pain.

Patients may experience adverse effects such as nausea, vomiting, somnolence, etc. when treated with opioids for moderately severe pain, but these adverse effects are generally tolerated by most patients within a week. However, constipation caused by opioid is not only high in occurrence rate of 90% -100%, but also does not develop tolerance due to long-term administration, and constipation occurs not only in the initial stage of administration but also continuously in the whole course of analgesic treatment. However, it is notable that constipation, if not controlled in time, can cause serious complications, which is the biggest obstacle to effective pain relief. Meanwhile, constipation can seriously affect the treatment of diseases, interrupt the treatment, greatly prolong the hospitalization time of patients and reduce the life quality of the patients. Therefore, the prevention and treatment of constipation adverse reactions is always a problem that opioid analgesic treatment periods cannot be ignored.

Research shows that opioid receptors exist in central nervous system (including brain and spinal cord), but also widely exist in peripheral nervous system in gastrointestinal tract and other organs, and opioid drugs combine with intestinal opioid receptors to slow intestinal peristalsis, reduce intestinal fluid secretion, increase intestinal fluid secretion, reduce activities of excitatory and inhibitory neurons in intestinal myolayer nerve plexus, increase muscular tension of intestinal wall smooth muscle and inhibit coordinated peristalsis, thereby increasing non-peristaltic contraction and finally causing constipation. Constipation will continue to exist during treatment due to the very slow development of intestinal tolerance to opioids in humans.

Drug therapy is a conventional means of relieving or treating constipation caused by opioids, and in particular, it is desirable to find opioids that block peripheral mu opioid receptors without affecting central analgesic effects. Opioid receptor antagonists do not themselves agonize opioid receptors, but have a strong affinity for mu receptors, and also have a certain affinity for kappa, delta and sigma receptors, and can remove opioid analgesics bound to these receptors, thereby producing an antagonistic effect. The systemic use of opioid receptor antagonists has effects on both central and peripheral opioid receptors, and antagonizes peripheral effects of opioids while also attenuating central analgesic effects such as naloxone, naltrexone, and nalmefene.

WO 2017/133634 A1 discloses a series of conjugates of polyethylene glycol and opioid receptor antagonists, and researches show that the double opioid receptor antagonist conjugates are more difficult to pass through the blood brain barrier and can better target the peripheral nervous system, so that the analgesic effect of opioid drugs is not affected while the toxic and side effects of the opioid drugs are better antagonized, and the double opioid receptor antagonist conjugates have wide clinical application prospect.

In pharmaceutical formulation and clinical application, it is desirable to use solid drugs with high stability. However, the dual opioid receptor antagonist conjugate is oily and viscous, and is difficult to be solid, and the need for further formulation development is not satisfied. There are no reports in the prior art on stable solid salt forms and polymorphs of the dual opioid receptor antagonist conjugate.

Disclosure of Invention

Problems to be solved by the invention

In order to ensure the physical and chemical stability of the medicine in the production and the medicine storage process, the solid salt and the polymorphic substance which can exist stably are screened out by deeply researching different acid addition salts of the double opioid receptor antagonist conjugate, and the corresponding preparation method is simple, convenient and easy to implement and has operability.

Solution for solving the problem

In a first aspect, the present invention provides a phosphate salt of a compound of formula I,

wherein the molar ratio of phosphoric acid to the compound of formula I is 1:1-3:1.

Preferably, in the phosphate of the compound of formula I, the molar ratio of phosphoric acid to the compound of formula I is 1.5:1 to 2.5:1.

More preferably, in the phosphate salt of the compound of formula I, the molar ratio of phosphoric acid to the compound of formula I is 2:1.

Further, the phosphate salt of the compound of formula I is amorphous.

Preferably, the X-ray powder diffraction (XRPD) pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 2.

In a second aspect, the invention provides a phosphate salt of the compound of formula I having a crystalline form 1, the XRPD pattern of which has characteristic peaks at the following 2Θ values: 4.3.+ -. 0.2 °, 5.1.+ -. 0.2 °, 6.2.+ -. 0.2 ° and 11.0.+ -. 0.2 °.

Preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 13.2±0.2°, 13.6±0.2°, 16.7±0.2°, 21.3±0.2° and 22.7±0.2°.

More preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 10.5±0.2°, 11.7±0.2°, 18.7±0.2°, 19.0±0.2°, 19.4±0.2° and 20.6±0.2°.

Further preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 12.

Further, the Differential Scanning Calorimetry (DSC) profile of the phosphate salt of the compound of formula I shows an endotherm at 191±1 ℃.

Preferably, the DSC profile of the phosphate salt of the compound of formula I is substantially in accordance with figure 9.

Further, the thermogravimetric analysis (TGA) profile of the phosphate salt of the compound of formula I shows a weight loss of about 0.05% at 179±1 ℃.

Preferably, the TGA profile of the phosphate salt of the compound of formula I is substantially in accordance with figure 9.

In a third aspect, the invention provides a phosphate salt of the compound of formula I having form 2 with an XRPD pattern having characteristic peaks at the following 2Θ values: 5.7 + -0.2 deg., 11.5 + -0.2 deg., 16.8 + -0.2 deg., and 17.1 + -0.2 deg..

Preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 3.8.+ -. 0.2 °, 4.9.+ -. 0.2 °, 6.7.+ -. 0.2 °, 11.1.+ -. 0.2 °, 13.6.+ -. 0.2 °, 14.0.+ -. 0.2 °, 19.5.+ -. 0.2 ° and 21.3.+ -. 0.2 °.

More preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 9.2±0.2°, 9.8±0.2°, 10.3±0.2°, 12.7±0.2°, 15.7±0.2°, 16.1±0.2° and 22.4±0.2°.

Further preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 11.

Further, the DSC profile of the phosphate salt of the compound of formula I shows endotherm at 119.+ -. 1 ℃, 184.+ -. 1 ℃ and 189.+ -. 1 ℃.

Preferably, the DSC profile of the phosphate salt of the compound of formula I substantially corresponds to figure 8.

Further, the TGA profile of the phosphate salt of the compound of formula I shows a weight loss of about 0.79% at 192+ -1deg.C.

Preferably, the TGA profile of the phosphate salt of the compound of formula I is substantially in accordance with FIG. 8.

In a fourth aspect, the present invention provides a phosphate salt of the compound of formula I having a crystalline form 3, the XRPD pattern of which has characteristic peaks at the following 2Θ values: 3.9 ± 0.2 °, 4.8 ± 0.2 °, 16.5 ± 0.2 ° and 16.7 ± 0.2 °.

Preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 7.2 + -0.2 deg. and 17.5 + -0.2 deg..

More preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 11.6 + -0.2 DEG and 14.2 + -0.2 deg.

Further preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 4.

Further, the DSC profile of the phosphate salt of the compound of formula I shows endotherm at 183+ -1deg.C and 189+ -1deg.C.

Preferably, the DSC profile of the phosphate salt of the compound of formula I substantially corresponds to figure 5.

Further, the TGA profile of the phosphate salt of the compound of formula I shows a weight loss of about 3.09% at 185+ -1deg.C.

Preferably, the TGA profile of the phosphate salt of the compound of formula I is substantially in accordance with FIG. 5.

In a fifth aspect, the invention provides a phosphate salt of the compound of formula I having a crystalline form 4 with an XRPD pattern having characteristic peaks at the following 2Θ values: 4.3 + -0.2 deg., 4.8 + -0.2 deg., and 16.5 + -0.2 deg..

Preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 12.3 ± 0.2 °.

More preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has characteristic peaks at the following 2θ values: 9.0.+ -. 0.2 °, 9.3.+ -. 0.2 ° and 17.6.+ -. 0.2 °.

Further preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 10.

Further, the DSC profile of the phosphate salt of the compound of formula I shows endotherms at 179+ -1deg.C and 187+ -1deg.C.

Preferably, the DSC profile of the phosphate salt of the compound of formula I is substantially in accordance with figure 7.

Further, the TGA spectrum of the phosphate salt of the compound of formula I shows a weight loss of about 1.51% at 189+ -1deg.C.

Preferably, the TGA profile of the phosphate salt of the compound of formula I is substantially in accordance with FIG. 7.

In a sixth aspect, the present invention provides a process for the preparation of a phosphate salt of a compound of formula I as described in the second aspect, selected from the group consisting of antisolvent processes and suspension-seeding processes, preferably antisolvent processes.

Preferably, the good solvent used in the anti-solvent method is water or a mixed solvent of fatty alcohol and water, preferably a mixed solvent of ethanol and water, and the anti-solvent used is fatty alcohol, preferably ethanol.

Further, the substance to be crystallized used in the antisolvent method is a phosphate of the compound of formula I according to any one of the first aspect and the third to fifth aspects.

Preferably, the solvent used in the suspension crystal transformation method is a mixed solvent of fatty alcohol and water, preferably a mixed solvent of ethanol and water, and more preferably an ethanol water solution with an ethanol-water volume ratio of 20:1-100:1.

Further, the substance to be crystallized used in the suspension-crystallization method is a phosphate of the compound of formula I as described in any one of the first aspect and the third to fifth aspects.

In a seventh aspect, the present invention provides a process for the preparation of a phosphate salt of a compound of formula I as described in the third aspect, selected from the group consisting of antisolvent processes and suspension-seeding processes, preferably antisolvent processes.

Preferably, the good solvent used in the anti-solvent method is a fatty alcohol, preferably methanol, and the anti-solvent used is a fatty alcohol, a fatty ether, a fatty ketone, a fatty acid ester, or any combination thereof, preferably ethanol-tetrahydrofuran, acetone or ethyl acetate, more preferably acetone.

Further, the substance to be crystallized used in the antisolvent method is a phosphate of the compound of formula I according to any one of the first to second aspects and the fourth to fifth aspects.

Preferably, the solvent used in the suspension crystal transformation method is water, fatty alcohol, fatty ketone or any combination thereof, and preferably ethanol water solution or acetone with the volume ratio of alcohol to water of 20:1.

Further, the substance to be crystallized used in the suspension-crystallization method is the phosphate of the compound of formula I described in the first or fourth aspect.

In an eighth aspect, the present invention provides a process for the preparation of a phosphate salt of a compound of formula I as described in the fourth aspect, which is a suspension-transcrystalline process.

Preferably, the solvent used in the suspension crystallization method is a fatty ether, a fatty ketone, a fatty acid ester, a fatty nitrile, or any combination thereof, preferably methyl tert-butyl ether-methyl formate, acetone or acetonitrile, more preferably methyl tert-butyl ether-methyl formate.

Further, the substance to be crystallized used in the suspension-crystallization method is the phosphate of the compound of formula I described in the first aspect.

In a ninth aspect, the present invention provides a process for the preparation of a phosphate salt of a compound of formula I as described in the fifth aspect, selected from the group consisting of antisolvent processes and suspension-seeding processes, preferably antisolvent processes.

Preferably, the good solvent used in the anti-solvent method is water, fatty alcohol or any combination thereof, preferably water or methanol, and the anti-solvent used is a fatty acid ester, preferably methyl formate.

Further, the substance to be crystallized used in the antisolvent method is a phosphate of the compound of formula I according to any one of the first to fourth aspects.

Preferably, the solvent used in the suspension-transfer method is a fatty alcohol, a fatty ether, a fatty acid ester, or any combination thereof, preferably tetrahydrofuran, methyl formate, or methyl-methanol-formate.

In a tenth aspect, the present invention provides a pharmaceutical composition comprising a phosphate salt of a compound of formula I according to any one of the first to fifth aspects.

Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

In an eleventh aspect, the present invention provides the use of a phosphate salt of a compound of formula I as defined in any one of the first to fifth aspects or a pharmaceutical composition in the tenth aspect for the manufacture of a medicament for the prevention and/or treatment of intestinal dysfunction caused at least in part by opioid.

In a twelfth aspect, the present invention provides a phosphate salt of a compound of formula I as described in any one of the first to fifth aspects or a pharmaceutical composition in the tenth aspect for use in the prevention and/or treatment of intestinal dysfunction caused at least in part by opioid.

In a thirteenth aspect, the present invention provides a method for preventing and/or treating bowel dysfunction caused at least in part by opioid comprising the steps of: administering a prophylactically and/or therapeutically effective amount of a phosphate salt of a compound of formula I according to any one of the first to fifth aspects or a pharmaceutical composition according to the tenth aspect to a subject in need thereof.

Further, the bowel dysfunction caused at least in part by the opioid is constipation.

ADVANTAGEOUS EFFECTS OF INVENTION

According to the invention, through screening of the acid addition salt of the opioid receptor antagonist conjugate, solid diphosphate and polymorphic substances thereof which can exist stably are obtained, wherein the crystal form 1 has excellent thermodynamic stability.

Drawings

FIG. 1 is a diagram of PEG8- (6-alpha-na Qu Mi) 2 free base ¹ H-NMR spectrum.

FIG. 2 is an XRPD pattern for the amorphous form of example 2.

FIG. 3 is a photograph of PLM of the amorphous material of example 2.

Fig. 4 is an XRPD pattern of the polymorph (form 3) produced in the methyl tert-butyl ether-methyl formate system in example 2.

FIG. 5 is a DSC and TGA overlay of the polymorph (form 3) prepared in example 2 in a methyl tert-butyl ether-methyl formate system.

Fig. 6 is an XRPD overlay pattern of two polymorphs obtained in example 2 at different acid-base molar ratios.

FIG. 7 is a DSC and TGA overlay of the polymorph (form 4) prepared in example 2 in a methyl methanol-formate system.

FIG. 8 is a DSC and TGA overlay of the polymorph (form 2) prepared in the methanol-acetone system of example 2.

FIG. 9 is a DSC and TGA overlay of the polymorph (form 1) prepared in the ethanol-water system in example 2.

Fig. 10 is an XRPD pattern of the polymorph (form 4) produced in the methyl methanol-formate system in example 2.

Fig. 11 is an XRPD pattern of the polymorph (form 2) produced in the methanol-ethanol-tetrahydrofuran system in example 2.

Fig. 12 is an XRPD pattern of the polymorph (form 1) produced in the ethanol-water system in example 2.

Fig. 13 is an XRPD overlay pattern of polymorphs produced in different systems in example 2.

Fig. 14 is an XRPD overlay pattern showing the results of a suspension seeding preliminary study in example 2.

Fig. 15 is an XRPD overlay pattern showing the results of the crystallization process optimization study in example 2.

Fig. 16 is an XRPD overlay pattern of four polymorphs produced using the optimization procedure of example 2.

Fig. 17 is an XRPD overlay pattern showing the seeding of polymorph (form 2) in a water-ethanol system (transition to form 1).

Fig. 18 is an XRPD overlay pattern showing the seeding of polymorph (form 3) in a water-ethanol system (transition from form 2 intermediate to form 1).

Fig. 19 is a DSC profile showing the seeding of polymorph (form 3) in a water-ethanol system (transition from form 2 intermediate to form 1).

Fig. 20 is an XRPD overlay pattern showing the seeding of polymorph (form 4) in a water-ethanol system (transition to form 1).

Fig. 21 is an XRPD overlay pattern showing the seeding situation (no seeding occurs) of a polymorph (form 1) in a methanol-acetone system.

Fig. 22 is an XRPD overlay pattern showing the seeding situation (neither seeding occurs) of the polymorph (form 1) under high temperature and mechanical milling conditions.

FIG. 23 is a DSC and TGA overlay of the polymorph (form 2) prepared in example 2 in a methanol-ethanol-tetrahydrofuran system.

Detailed Description

Unless otherwise defined, scientific and technical terms herein have the meanings commonly understood by one of ordinary skill in the art.

Unless otherwise indicated, the singular forms herein, such as "a," "an," and "the," encompass the plural referents thereof unless the context clearly dictates otherwise. For example, when reference is made to "a" crystal or polymorph, one or more different crystals or polymorphs are contemplated, and when reference is made to "the" method, equivalent steps and methods known to those of ordinary skill in the art are contemplated.

Unless otherwise indicated, the terms "comprises" and variations such as "comprising" and "includes" appearing herein mean that the collection encompasses not only the explicitly disclosed integer or integers, steps or combination thereof, but also does not exclude any other integer or step or combination thereof. Meanwhile, the term "comprising" appearing herein may be replaced by the term "containing", "including" or "having" in certain cases.

For the crystalline forms herein, only the characteristic peaks (i.e., the most characteristic, significant, unique and/or repeatable diffraction peaks) in the XRPD pattern are summarized, while other peaks may be obtained from the pattern by conventional methods. The characteristic peaks described above may be repeated within the margin of error (error range of + -0.2 deg.).

Solid salts of opioid receptor antagonist conjugates

In view of the problems of the opioid receptor antagonist conjugate being highly viscous, difficult to cure, and unable to meet formulation requirements, the present invention contemplates that solids suitable for formulation can be obtained by forming an acid addition salt with an acid. Through a number of salt and crystal form screening experiments, the present invention successfully discovers phosphates (especially bisphosphates) which can exist stably in solid form (especially in crystal form).

The present invention provides solid salts of opioid receptor antagonist conjugates wherein the free base moiety may be PEG8- (6- α -na Qu Mi) 2 (having a structure as shown in formula I) and the acid moiety may be phosphoric acid.

In one embodiment, the solid salt of the above opioid receptor antagonist conjugate is a phosphate salt of PEG8- (6- α -na Qu Mi) 2, wherein the molar ratio of phosphoric acid to free base can be 1:1 to 3:1, such as 1:1, 1.1:1, 1.3:1, 1.5:1, 1.7:1, 1.9:1, 2:1, 2.2:1, 2.4:1, 2.6:1, 2.8:1, 3:1, or any ratio in the above ranges.

In one embodiment, the molar ratio of phosphoric acid to free base in the solid salt of the above opioid receptor antagonist conjugate may be 1.5:1 to 2.5:1, e.g., 1:5, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, 2.5:1, or any ratio in the above ranges.

In one embodiment, the molar ratio of phosphoric acid to free base in the solid salt of the above opioid receptor antagonist conjugate may be 2:1.

In one embodiment, the solid salt of the opioid receptor antagonist conjugate described above is a phosphate salt of PEG8- (6- α -na Qu Mi) 2, which may be present in the form of an amorphous form. Under certain conditions, the XRPD pattern of the amorphous form may be substantially in accordance with figure 2.

In another embodiment, the solid salt of the above opioid receptor antagonist conjugate is a phosphate salt of PEG8- (6- α -na Qu Mi) 2, which may exist in the form of a polymorph. The polymorphs can have a variety of forms and can be characterized by at least one of XRPD, DSC, and TGA methods.

In one embodiment, the polymorph described above can have form 1. The XRPD pattern of the polymorph having form 1 can have characteristic peaks at the following 2θ values: 4.3±0.2°, 5.1±0.2°, 6.2±0.2° and 11.0±0.2°, it is preferable that the characteristic peaks at the following 2θ values may also be present: 13.2±0.2°, 13.6±0.2°, 16.7±0.2°, 21.3±0.2° and 22.7±0.2°, more preferably, it is also possible to have characteristic peaks at the following 2θ values: 10.5±0.2°, 11.7±0.2°, 18.7±0.2°, 19.0±0.2°, 19.4±0.2° and 20.6±0.2°, and more preferably, may substantially correspond to fig. 12.

And/or the DSC profile of the polymorph with form 1 can exhibit an endotherm at 191±1 ℃, preferably can be substantially in accordance with fig. 9.

And/or the TGA profile of the polymorph having form 1 can exhibit a weight loss of about 0.05% at 179±1 ℃, preferably can be substantially in accordance with fig. 9.

In another embodiment, the polymorph described above can have form 2. The XRPD pattern of the polymorph having form 2 can have characteristic peaks at the following 2θ values: 5.7±0.2°, 11.5±0.2°, 16.8±0.2° and 17.1±0.2°, it being preferable that the characteristic peaks at the following 2θ values are also present: 3.8±0.2°, 4.9±0.2°, 6.7±0.2°, 11.1±0.2°, 13.6±0.2°, 14.0±0.2°, 19.5±0.2° and 21.3±0.2°, and more preferably, can also have characteristic peaks at the following 2θ values: 9.2.+ -. 0.2 °, 9.8.+ -. 0.2 °, 10.3.+ -. 0.2 °, 12.7.+ -. 0.2 °, 15.7.+ -. 0.2 °, 16.1.+ -. 0.2 ° and 22.4.+ -. 0.2 °, further preferably may substantially correspond to fig. 11.

And/or the DSC profile of the polymorph with form 2 can exhibit endotherms at 119±1 ℃, 184±1 ℃ and 189±1 ℃, preferably can be substantially in accordance with fig. 8.

And/or the TGA profile of the polymorph having form 2 can exhibit a weight loss of about 0.79% at 192±1 ℃, preferably can be substantially in accordance with fig. 8.

In yet another embodiment, the polymorph described above can have form 3. The XRPD pattern of the polymorph having form 3 can have characteristic peaks at the following 2θ values: 3.9±0.2°, 4.8±0.2°, 16.5±0.2° and 16.7±0.2°, preferably may also have characteristic peaks at the following 2θ values: 7.2±0.2° and 17.5±0.2°, more preferably, may also have characteristic peaks at the following 2θ values: 11.6 + 0.2 deg. and 14.2 + 0.2 deg., and more preferably may be substantially identical to fig. 4.

And/or the DSC profile of the polymorph with form 3 can exhibit endotherms at 183±1 ℃ and 189±1 ℃, preferably can be substantially in accordance with figure 5.

And/or the TGA profile of the polymorph having form 3 can exhibit a weight loss of about 3.09% at 185±1 ℃, preferably can be substantially in accordance with fig. 5.

In yet another embodiment, the polymorph described above can have form 4. The XRPD pattern of the polymorph having form 4 can have characteristic peaks at the following 2θ values: 4.3±0.2°, 4.8±0.2° and 16.5±0.2°, preferably may also have characteristic peaks at the following 2θ values: 12.3±0.2°, more preferably may also have characteristic peaks at the following 2θ values: 9.0±0.2°, 9.3±0.2° and 17.6±0.2°, and more preferably may substantially correspond to fig. 10.

And/or the DSC profile of the polymorph with form 4 can exhibit endotherms at 179±1 ℃ and 187±1 ℃, preferably can be substantially in accordance with fig. 7.

And/or the TGA profile of the polymorph having form 4 can exhibit a weight loss of about 1.51% at 189±1 ℃, preferably can be substantially in accordance with fig. 7.

Process for preparing solid salts of opioid receptor antagonist conjugates

The present invention also provides various methods of preparing solid salts of the above opioid receptor antagonist conjugates (e.g., the diphosphate salt of PEG8- (6- α -na Qu Mi) 2).

In one embodiment, the diphosphate of PEG8- (6- α -na Qu Mi) 2 described above can be prepared by the following method: respectively dissolving phosphoric acid and PEG8- (6-alpha-Na Qu Mi) 2 in a solvent to obtain an acid solution and a free alkali solution; and mixing and stirring the acid solution and the free alkali according to a proportion, and desolventizing to obtain the diphosphate of PEG8- (6-alpha-Na Qu Mi) 2.

In one embodiment, the solvent in the above method may be an organic solvent, preferably may be a fatty alcohol, a fatty ether or a fatty acid ester, more preferably may be methanol, tetrahydrofuran or methyl formate, and still more preferably may be methanol.

The concentration of the acid solution and/or the free alkali solution in the above-mentioned method is not particularly limited in any way, and for example, the concentration of the acid solution may be 0.01 to 1mol/L (particularly 0.1 mol/L).

In one embodiment, the acid-base ratio in the above method may be a molar ratio, preferably an acid-base molar ratio of 1:1 to 3:1, more preferably an acid-base molar ratio of 1:1 or 2:1, and still more preferably an acid-base molar ratio of 2:1.

The stirring conditions (e.g., temperature, time period, etc.) in the above-described method are not subject to any specific limitation, and for example, stirring may be carried out at room temperature for 15 minutes.

The present invention is not limited in any particular way with respect to the desolventizing conditions (e.g., temperature, pressure, duration, etc.) in the above-described method, and may be carried out, for example, by means of reduced pressure distillation (e.g., using a rotary evaporator).

In one embodiment, the diphosphate salt of PEG8- (6- α -na Qu Mi) 2 in the above method may be an off-white solid.

In the present invention, the biphosphate of PEG8- (6-alpha-na Qu Mi) 2 prepared by the above method may exist in the form of an amorphous substance, and its specific characterization method and results are described above.

In addition, the invention also provides a preparation method of the biphosphate of PEG8- (6-alpha-Na Qu Mi) 2 in the form of a polymorph.

First, the polymorph of PEG8- (6- α -na Qu Mi) 2 diphosphate has a crystal form 1, and the preparation method may be an antisolvent method or a suspension crystal transformation method, and preferably may be an antisolvent method.

In one embodiment, the good solvent used in the above-mentioned anti-solvent method may be water or a mixed solvent of fatty alcohol and water, and the anti-solvent used may be fatty alcohol.

In one embodiment, the good solvent used in the above-mentioned anti-solvent method may be a mixed solvent of ethanol and water, and the anti-solvent used may be ethanol.

In one embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6-alpha-na Qu Mi) 2 in the form of an amorphous substance.

In another embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 2.

In yet another embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 3.

In yet another embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 4.

In one embodiment, the solvent used in the suspension crystallization method may be a mixed solvent of fatty alcohol and water, preferably a mixed solvent of ethanol and water, and more preferably an aqueous ethanol solution having an aqueous alcohol-to-water ratio of 20:1 to 100:1.

In one embodiment, the substance to be crystallized used in the above suspension crystallization method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of an amorphous substance.

In another embodiment, the material to be crystallized used in the above suspension crystallization method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 2.

In yet another embodiment, the material to be crystallized used in the above suspension crystallization method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 3.

In yet another embodiment, the material to be crystallized used in the above suspension crystallization method may be a diphosphate salt of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 4.

Secondly, the polymorph of the biphosphate of PEG8- (6-alpha-nano Qu Mi) 2 has a crystal form 2, and the preparation method can be an antisolvent method or a suspension crystal transformation method, and preferably can be an antisolvent method.

In one embodiment, the good solvent used in the above anti-solvent method may be a fatty alcohol, and the anti-solvent used may be a fatty alcohol, a fatty ether, a fatty ketone, a fatty acid ester, or any combination thereof.

In one embodiment, the good solvent used in the above anti-solvent method may be methanol, and the anti-solvent used may be ethanol-tetrahydrofuran, acetone or ethyl acetate, and preferably may be acetone.

In another embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 1.

In one embodiment, the solvent used in the suspension crystallization method may be water, aliphatic alcohol, aliphatic ketone or any combination thereof, and preferably an ethanol water solution or acetone with an alcohol-water volume ratio of 20:1.

In another embodiment, the material to be crystallized used in the above suspension crystallization method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 3.

Thirdly, the polymorph of the biphosphate of PEG8- (6-alpha-Na Qu Mi) 2 has a crystal form 3, and the preparation method can be a suspension crystal transformation method.

In one embodiment, the solvent used in the suspension crystallization method may be a fatty ether, a fatty ketone, a fatty acid ester, a fatty nitrile, or any combination thereof, and preferably may be methyl tert-butyl ether-methyl formate, acetone or acetonitrile, and more preferably may be methyl tert-butyl ether-methyl formate.

Fourth, the polymorph of PEG8- (6- α -na Qu Mi) 2 diphosphate has form 4, and the preparation method may be an antisolvent method or a suspension crystal transformation method, and preferably may be an antisolvent method.

In one embodiment, the good solvent used in the above anti-solvent method may be water, a fatty alcohol, or any combination thereof, and the anti-solvent used may be a fatty acid ester.

In one embodiment, the good solvent used in the above-described anti-solvent method may be water or methanol, preferably methanol, and the anti-solvent used may be methyl formate.

In yet another embodiment, the substance to be crystallized used in the above anti-solvent method may be a diphosphate of PEG8- (6- α -na Qu Mi) 2 in the form of a polymorph having form 2.

In one embodiment, the solvent used in the suspension crystallization method may be a fatty alcohol, a fatty ether, a fatty acid ester, or any combination thereof, and preferably may be tetrahydrofuran, methyl formate, or methyl-methanol-formate.

Pharmaceutical composition comprising solid salts of opioid receptor antagonist conjugates

In the present invention, the solid salts of the above opioid receptor antagonist conjugates may be used either alone or in combination with other substances, and thus the present invention also provides a pharmaceutical composition which may comprise the solid salts of the opioid receptor antagonist conjugates of the present invention in any form, particularly in the form of an amorphous form or a polymorph having any of forms 1 to 4.

In one embodiment, the above pharmaceutical composition may further comprise at least one pharmaceutically acceptable excipient having meaning well known in the art.

Medical use of solid salts of opioid receptor antagonist conjugates or pharmaceutical compositions comprising same

In the present invention, the solid salts of opioid receptor antagonist conjugates, whether in any form (particularly in the form of an amorphous form or a polymorph having any of forms 1-4), or pharmaceutical compositions comprising the solid salts, are useful for preventing and/or treating bowel dysfunction caused at least in part by opioid drugs.

Thus, in one aspect, the present invention provides the use of a solid salt of an opioid receptor antagonist conjugate or a pharmaceutical composition comprising the same, in any form, particularly in the form of an amorphous form or a polymorph having any one of forms 1 to 4, for the manufacture of a medicament for the prevention and/or treatment of intestinal dysfunction caused at least in part by an opioid. In another aspect, the present invention also provides a method for preventing and/or treating bowel dysfunction caused at least in part by opioid, which may include the steps of: a prophylactically and/or therapeutically effective amount of a solid salt of an opioid receptor antagonist conjugate in any form (particularly in the form of an amorphous or polymorph having any one of forms 1-4) or a pharmaceutical composition comprising the same is administered to a subject in need thereof.

In one embodiment, the above-described bowel dysfunction caused at least in part by opioid may be constipation.

The invention will be further illustrated by the following specific examples. The instruments, practices, materials, and the like used in the following examples are all available through conventional commercial means unless otherwise indicated.

Analytical method and apparatus

(1) Nuclear magnetic resonance hydrogen spectrum [ ] ¹ H-NMR)

¹ The H-NMR spectrum was acquired using a Bruker Advance 300M NMR apparatus equipped with a B-ACS 120 autosample injection system.

(2) X-ray powder diffraction (XRPD)

X-ray powder diffraction A Bruker D8 advanced powder X-ray diffractometer and a Bruker D2 phaser powder X-ray diffractometer equipped with Lynxeye detector were used. The Bruker D8 advanced powder X-ray diffractometer is adopted to test the sample, the 2 theta scanning angle is from 3 degrees to 40 degrees, the scanning step length is 0.02 degrees, and the light tube voltage and the light tube current when the sample is measured are 40kV and 40mA respectively. The 2 theta scanning angle of the sample is 3-40 degrees, the scanning step length is 0.02 degrees, and the light tube voltage and the light tube current are 30kV and 10mA respectively when the sample is measured by using a Bruker D2 phaser powder X-ray diffractometer.

(3) Polarized light microscopic analysis (PLM)

PLM analysis was performed using a Nikon Eclipse LV100N POL polarization microscope.

(4) Thermogravimetric analysis (TGA)

Thermogravimetric analysis employed a TGA Q500 thermogravimetric analyzer or a Discovery TGA 55 thermogravimetric analyzer. The test samples were placed in equilibrated open aluminum sample trays and the mass was automatically weighed in a TGA oven. The sample was heated to the final temperature at a ramp rate of 10 c/min.

(5) Differential scanning calorimetric analysis (DSC)

Differential scanning calorimetric analysis employed a DSC Q200 differential scanning calorimeter or Discovery DSC 250 differential scanning calorimeter. After accurate weighing, the test samples were placed in DSC-punch sample trays and the exact mass of the samples was recorded. The sample was heated to the final temperature at a ramp rate of 10 c/min.

(6) Ion Chromatography (IC)

The ion chromatography adopts a Thermo Fischer ICS-5000+ ion chromatography system, and the chromatographic column is IonPac ^TM AS11 (4X 250 mM), mobile phase was 30mM KOH aqueous solution, flow rate 1.0mL/min. Accurately weighing 25+/-2 mg of sample into a 100mL volumetric flask, adding 2/3 of the volume of water for dissolution (ultrasonic when necessary), then using the water for volume fixing, and uniformly mixing to obtain a sample solution. The chromatographic system and chromatographic column are balanced to a base line by using a mobile phase balance, and the content of phosphate ions in a sample is calculated according to the following formula by sequentially sampling a blank solution (water), a quantitative limiting solution (the concentration is 4 mug/mL based on phosphate ions), a working standard solution (the concentration is 40 mug/mL based on phosphate ions) and repeated sampling for 6 times), a retest standard solution (the concentration is the same as the working standard solution), the blank solution, a sample solution and the working standard solution:

A (spl): peak area of phosphate ions in the sample solution;

a (std): average value of phosphate ion peak area in initial six-needle continuous working standard solution;

conco (std): phosphate ion concentration in phosphate ion standard solution (1000. Mu.g/mL);

wt (spl): mass of sample (mg);

DF (spl): a dilution factor (100) of the sample;

DF (std): dilution factor of working standard (25).

Solvents used and abbreviations therefor

Methanol (MeOH), toluene, methyl tert-butyl ether (MTBE), methyl Ethyl Ketone (MEK), methyl Formate (MF), acetone, ethanol (EtOH), acetonitrile (ACN), ethyl Acetate (EA), dichloromethane (DCM), tetrahydrofuran (THF), isopropyl alcohol (IPA), water (W).

Example 1: salt screening of opioid receptor antagonist conjugates

1.96 well plate salification screening

14 acids (hydrochloric acid, hydrobromic acid, sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, maleic acid, phosphoric acid, boric acid, palmitic acid, fumaric acid, citric acid, succinic acid and L-tartaric acid) were dissolved in methanol, respectively, to prepare 0.1mol/L acid solutions.

PEG8- (6-alpha-na Qu Mi) 2 free base (prepared according to the method described in WO 2017/133634 A1) ¹ H-NMR spectrum as shown in FIG. 1) was dissolved in methanol to prepare a 30mg/mL solution, which was plated on a 96-well plate, three plates were plated in parallel, and 100. Mu.L of each well was added.

Hydrochloric acid, hydrobromic acid, sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, maleic acid and phosphoric acid were used as acid A, and two groups were arranged in parallel, the first group adding 30. Mu.L/well of an acid solution to a first 96-well plate so that the molar ratio of acid A to free base was about 1:1, and the second group adding 60. Mu.L/well of an acid solution to a second 96-well plate so that the molar ratio of acid A to free base was about 2:1. With boric acid, palmitic acid, fumaric acid, citric acid, succinic acid and L-tartaric acid as acid B, 60. Mu.L/well of acid solution was added to the third 96-well plate such that the molar ratio of acid B to free base was about 2:1.

After methanol was completely volatilized, 200. Mu.L of solvent was added to each well, and the mixture was sealed with a sealing film, and placed in a fume hood at room temperature to evaporate the solvent. The specific results are shown in Table 1.

TABLE 1 results of salt formation screening of opioid receptor antagonist conjugate free base with various acids in various solvents

1

2

3

4

5

6

7

8

Hydrochloric acid

Hydrobromic acid

Sulfuric acid

Para-toluene sulfonic acid

Methanesulfonic acid

Oxalic acid

Maleic acid

Phosphoric acid

1

Toluene (toluene)

Oil (oil)

2

MTBE

Oil (oil)

3

MEK

Oil (oil)

4

MF

Oil (oil)

Solid body

5

Acetone (acetone)

Oil (oil)

6

EtOH

Oil (oil)

7

ACN

Oil (oil)

8

EA

Oil (oil)

9

DCM

Oil (oil)

10

THF

Oil (oil)

Solid body

11

IPA

Oil (oil)

Oil |

Oil (oil)

12

Water and its preparation method

Oil (oil)

A

B

C

D

E

F

Boric acid

Palmitic acid

Fumaric acid

Citric acid

Isonicotinic acid

L-tartaric acid

1

Toluene (toluene)

Oil (oil)

2

MTBE

Oil (oil)

Oil/solid

Oil (oil)

3

MEK

Oil (oil)

4

MF

Oil (oil)

Oil/solid

Oil (oil)

5

Acetone (acetone)

Oil (oil)

Solid body

6

EtOH

Oil (oil)

Oil/solid

Oil (oil)

7

ACN

Oil (oil)

Solid body

8

EA

Oil (oil)

9

DCM

Oil (oil)

10

THF

Oil (oil)

11

IPA

Oil (oil)

12

Water and its preparation method

Oil (oil)

The results show that after evaporation of the solvent in the 96-well plate, the samples in the wells are mostly glassy oils. Wherein phosphate is solid in MF and THF; palmitate gives a mixture of oil and solids in MTBE, MF, etOH; the L-tartrate is solid in acetone and ACN, but is oily when repeated experiments are carried out, so that the process reproducibility is low; the remaining samples were all oily.

2. Salt formation studies of phosphoric acid and free base under different ratio conditions

A methanol solution containing 0.1mol/L phosphoric acid was added to a methanol solution containing a free base (molar ratio of acid to base: 1:1 and 2:1, respectively), and the mixture was stirred at room temperature for 15 minutes and then dried by spin to give an off-white solid. Adding a mixture solution of ethanol and water (the ratio of the ethanol to the water is about 150:1), heating to 45 ℃ to dissolve, naturally cooling to room temperature, and stirring overnight to precipitate a white solid. Characterization by XRPD (as shown in fig. 6), results show that the same crystal (form 1) is obtained at two acid-base ratios; the content of phosphate ions in both crystals was about 15.6wt% as determined by ion chromatography, indicating that the free base and phosphoric acid formed an acid addition salt at a 1:2 molar ratio.

Example 2: crystal form screening of opioid receptor antagonist conjugate phosphate

1. Preparation and characterization of the starting Material

This example prepared phosphate in both crystalline and amorphous forms as starting material for crystalline form screening.

Phosphate amorphous material: the reaction of PEG8- (6- α -na Qu Mi) 2 free base (prepared according to the method described in WO 2017/133634 A1) with phosphoric acid in methanol (molar ratio of acid to free base about 1:1) gives a methanolic solution of phosphate, which is then evaporated to dryness by rotary evaporator and characterized by XRPD (as shown in fig. 2) and PLM (as shown in fig. 3) with no apparent diffraction peaks in the pattern, no apparent crystalline particles in the photograph, and the character of the amorphous form.

Phosphate polymorphs: the phosphate amorphous prepared above was suspended in the MTBE-MF system and stirred overnight to give the phosphate polymorph (form 3) and characterized by XRPD, DSC and TGA, the results are shown in table 2 and fig. 4-5.

TABLE 2 XRPD characterization results for form 3

Angle (°)	d value	Relative intensity (%)
			3.851	22.92738	61.50％
4.793	18.42019	100.00％
			7.221	12.23261	15.10％
8.846	9.98822	8.10％
			8.997	9.82073	6.80％
10.725	8.24233	7.10％
			11.621	7.60870	13.00％
14.151	6.25346	11.00％
			16.492	5.37093	43.50％
16.653	5.31938	55.60％
			17.534	5.05404	25.70％
29.997	2.97650	5.00％

2. Early stage crystal form screening

For the amorphous or polymorphic forms of phosphate prepared in item 1, parallel experiments were performed with MTBE, meOH and water as good solvents and MF, THF and EtOH as anti-solvents, respectively, yielding 4 different crystalline salts (phosphate polymorphs), the specific preparation methods being shown in table 3.

TABLE 3 preparation of crystalline salts of opioid receptor antagonist conjugates

As can be seen from fig. 9 (form 1), the TGA profile of the crystalline salt obtained in the ethanol-water system shows little weight loss (0.05%) before decomposition, and the DSC profile shows an endothermic peak as a single peak, with the result being superior to the crystalline salt obtained in the other two systems. As can be seen from fig. 7 (form 4) and fig. 23 (form 2), TGA spectra of the crystalline salt obtained in the other two systems show more weight loss, and DSC spectra show no endothermic peak, which indicates that the conditions such as crystal transformation or mixed crystal may occur during the heating process. The crystallinity of the products obtained with other good solvent-antisolvent systems is poor, essentially as a mixture of amorphous and crystalline materials or as an oil, which is not described in detail herein.

The three crystalline salts obtained in the MeOH-MF, meOH-EtOH-THF and EtOH-water systems were characterized by XRPD and the results are shown in tables 4-6 and FIGS. 10-12, respectively.

TABLE 4 XRPD characterization results for form 4

Angle (°)	d value	Relative intensity (%)
			4.295	20.55706	100.00％
4.763	18.53745	79.70％
			7.814	11.30512	5.70％
8.989	9.82977	12.00％
			9.261	9.54126	10.40％
11.098	7.96588	7.70％
			11.675	7.57350	8.80％
12.311	7.18409	16.60％
			14.968	5.91409	4.50％
15.136	5.84884	7.80％
			16.478	5.37540	52.30％
17.622	5.02878	13.60％
			23.277	3.81831	5.00％

TABLE 5 XRPD characterization results for Crystal form 2

Angle (°)	d value	Relative intensity (%)
			3.781	23.35182	28.10％
4.876	18.10748	23.10％
			5.682	15.54054	100.00％
6.697	13.18724	16.60％
			9.214	9.59060	14.60％
9.817	9.00285	11.80％
			10.313	8.57051	12.70％
11.077	7.98145	20.20％
			11.504	7.68600	58.80％
12.680	6.97543	14.70％
			13.615	6.49867	22.10％
13.996	6.32261	19.60％
			15.661	5.65382	13.60％
16.119	5.49418	11.80％
			16.759	5.28598	49.30％
17.123	5.17423	35.60％
			18.674	4.74800	5.20％
19.540	4.53940	16.10％
			20.766	4.27409	6.20％
21.061	4.21476	7.80％
			21.313	4.16556	16.10％
22.368	3.97149	12.50％
			22.778	3.90086	7.10％
23.909	3.71879	3.20％
			25.755	3.45638	4.20％
26.384	3.37534	5.40％
			28.619	3.11659	2.50％
29.446	3.03088	4.50％

TABLE 6 XRPD characterization results for Crystal form 1

Angle (°)	d value	Relative intensity (%)
			4.336	20.36359	100.00％
5.136	17.19232	53.20％
			6.240	14.15254	69.90％
7.042	12.54352	7.90％
			7.571	11.66756	3.70％
9.018	9.79813	5.00％
			10.127	8.72748	8.00％
10.524	8.39957	14.50％
			10.965	8.06236	39.10％
11.655	7.58649	12.20％
			12.611	7.01367	5.30％
13.220	6.69162	26.40％
			13.552	6.52880	23.80％
14.484	6.11077	2.40％
			14.895	5.94303	5.00％
15.471	5.72290	7.50％
			16.092	5.50330	2.20％
16.704	5.30325	22.60％
			17.051	5.19604	3.50％
17.630	5.02656	4.10％
			18.157	4.88178	3.90％
18.652	4.75343	12.50％
			19.047	4.65565	11.80％
19.373	4.57802	11.20％
			20.301	4.37094	2.60％
20.631	4.30181	10.20％
			21.253	4.17716	15.00％
22.215	3.99839	5.70％
			22.729	3.90909	17.90％
23.313	3.81259	9.20％
			23.575	3.77070	7.20％
24.638	3.61046	4.50％
			25.273	3.52117	3.90％
26.445	3.36772	3.50％
			26.952	3.30551	1.50％
27.992	3.18503	3.90％
			28.810	3.09635	1.80％
30.028	2.97352	1.50％
			31.022	2.88043	3.00％
34.156	2.62297	1.80％

From the above results and fig. 13, it is understood that the 4 kinds of crystal salts (phosphate polymorphs) obtained by the above method have different crystal forms.

3. Suspension transcrystalline preliminary study (conversion of amorphous to polymorph)

The single solvent suspension crystal transformation research is carried out by using the phosphate amorphous substance prepared in the 1 st step, and the specific method is as follows: about 200mg of the sample was weighed, 3mL of solvent was added, and the mixture was stirred at room temperature for 2 days, and after sampling, XRPD analysis was performed, the results of which are shown in FIG. 14. The results show that after suspension seeding in ethanol, ethyl acetate, methylene chloride, butanone, isopropanol, methyl tertiary butyl ether and toluene, the phosphate remains amorphous or gives a product that is approximately amorphous; after suspension crystal transformation in tetrahydrofuran and methyl formate, the same new phosphate polymorph (crystal form 4) is obtained; after suspension seeding in acetone, another new polymorph (form 2) is obtained, but with lower crystallinity; after suspension seeding in acetonitrile, the phosphate polymorph prepared in item 1 (form 3) is obtained, but the crystallinity is too low.

4. Optimization research of crystallization process

Because of the flexible PEG chain in the structure of PEG8- (6- α -na Qu Mi) 2 free base, the phosphate is more difficult to crystallize, and only amorphous or low crystallinity polymorphs can be obtained under most conditions in item 3. Therefore, based on the above-described results, the amorphous or crystalline phosphate prepared in item 1 was used as a starting material, an appropriate good solvent-antisolvent system was selected to optimize the crystallization process, and finally methanol and water were used as good solvents, and methyl formate, ethanol, tetrahydrofuran, acetone, ethyl acetate, and methylene chloride were used as antisolvents, as shown in table 7.

TABLE 7 optimized preparation Process of crystalline salts of opioid receptor antagonist conjugates

As can be seen from fig. 15, polymorphs having form 1 can be obtained in ethanol-water systems and have better crystallinity; polymorphs having form 4 can be obtained in both the methyl formate-water system and the methyl formate-methanol system, but with relatively low crystallinity; polymorphs with form 2 can be obtained in methanol-acetone system, methanol-ethyl acetate system and methanol-ethanol-tetrahydrofuran system, and have better crystallinity; whereas in the methanol-dichloromethane system only amorphous or near amorphous products are obtained.

The optimal process for preparing 4 phosphate polymorphs having different crystal forms was finally determined:

a sample of phosphate (e.g., 61.2mg of the polymorph of phosphate prepared in item 1 (form 3)) was weighed, water and ethanol (e.g., 0.06mL of water, 1mL of ethanol) were added sequentially, an oil started to appear, then heated to 40 ℃ and stirred (e.g., 3 hours), then slowly cooled to room temperature and stirred (e.g., 24 hours), and suction filtered to give white crystals (form 1).

A sample of phosphate (e.g., 197.6mg of amorphous phosphate prepared in item 1) was weighed, dissolved by adding methanol (e.g., 0.4 mL), then slowly added acetone (e.g., 4 mL), the solid was precipitated, stirred at room temperature (e.g., overnight), and suction filtered to give white crystals (form 2).

A certain amount of phosphate sample (for example, phosphate amorphous prepared by weighing 8.8g of free alkali methanol solution (14.47 wt%) and adding 14mL of 0.1mol/L phosphoric acid methanol solution, reacting for 15min, spin-drying the solvent to obtain the product), adding a mixed solution of methyl formate and methyl tert-butyl ether (for example, 2mL of methyl formate and 10mL of methyl tert-butyl ether), stirring at room temperature (for example, overnight), and suction-filtering to obtain white crystals (crystal form 3).

A sample of phosphate (e.g., phosphate amorphous prepared by weighing 3.5g of a free base methanol solution (14.47 wt%) and adding 5.2mL of a 0.1mol/L methanol phosphate solution, reacting for 30min, spin-drying the solvent to obtain the product), adding methanol (e.g., 0.5 mL), adding methyl formate (e.g., 7.5 mL) after an oil appears, converting the oil into a solid, adding methyl formate (e.g., 5 mL), stirring at room temperature (e.g., 1 hr), and suction-filtering to obtain white crystals (form 4).

XRPD analysis was performed on the four polymorphs described above, each having forms 1-4, with the results shown in fig. 16. From the XRPD pattern, forms 3 and 4 exhibited lower crystallinity, and forms 1 and 2 exhibited better crystallinity.

5. Suspension transcrystalline in depth (conversion between different polymorphs)

(1) Crystal form 2 crystal transformation in ethanol-water system

A sample of the phosphate polymorph (form 2) was added to a 1% water ethanol system and stirred at 40 ℃ for 2 days to give a solid sample that showed complete conversion to form 1 (as shown in figure 17).

(2) Crystal form 3 crystal transformation in ethanol-water system

About 300mg of the phosphate polymorph (form 3) was taken, a mixed solution of ethanol and water (containing 0.15ml of water and 3ml of ethanol) was added, and stirred at room temperature for 4 hours, and XRPD detection showed a change in form; at 40 ℃ to 8 hours, XRPD detection showed complete conversion to form 2; by 12 hours, form 2 began to transition to form 1, and by 30 hours XRPD detection showed complete transition to form 1 (as shown in fig. 18).

DSC results also reflect the conversion process from form 3 to form 1, wherein a gradual conversion from two endothermic peaks to one endothermic peak is shown (as shown in figure 19).

(3) Crystal form 4 crystal transformation in ethanol-water system

A sample of the phosphate polymorph (form 4) was added to a 1% water ethanol system and stirred at 40 ℃ for 2 days to give a solid sample that showed complete conversion to form 1 (as shown in figure 20).

(4) Crystal form 1 crystal transformation study in methanol-acetone system

The above results show that forms 2-4 can be converted into form 1 under certain conditions, and the preliminary results show that form 1 may be the dominant form. Since the specific type of the solvent may affect the transformation of the crystal form, a suspension crystal transformation mode is adopted to further examine whether the crystal form 1 is transformed in other systems.

The sample of the phosphate polymorph (form 1) was added to a methanol-acetone system for suspension crystallization, slurried and stirred for 2 days to give a solid sample, which showed no change in XRPD detection (as shown in fig. 21). The results show that even though form 1 is subjected to suspension seeding under the process conditions for preparing form 2, it cannot be converted into form 2, and the stability of form 1 is relatively high.

6. Stability study of Crystal form 1 under high temperature and mechanical grinding conditions

High temperature stability study: samples of the phosphate polymorph (form 1) were baked in an oven at 65 ℃ for 3 days and sampled for XRPD detection.

Mechanical grinding stability; samples of the phosphate polymorph (form 1) were mechanically milled for 3 minutes and XRPD detection was performed after sampling.

XRPD detection also showed no change (as shown in fig. 22), further indicating that form 1 has higher stability, suitable for subsequent formulation development.

Claims

1. The phosphate salt of the compound of formula I,

wherein the molar ratio of phosphoric acid to the compound of formula I is from 1:1 to 3:1, preferably from 1.5:1 to 2.5:1, more preferably 2:1.

2. A phosphate of a compound of formula I according to claim 1,

the phosphate of the compound of formula I is amorphous;

preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially in accordance with figure 2.

3. A phosphate of a compound of formula I according to claim 1,

the phosphate of the compound of formula I has form 1 and satisfies at least one of the conditions I to III:

condition I:

its XRPD pattern has characteristic peaks at the following 2θ values: 4.3±0.2°, 5.1±0.2°, 6.2±0.2° and 11.0±0.2°;

Preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 13.2±0.2°, 13.6±0.2°, 16.7±0.2°, 21.3±0.2° and 22.7±0.2°;

more preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 10.5±0.2°, 11.7±0.2°, 18.7±0.2°, 19.0±0.2°, 19.4±0.2° and 20.6±0.2°;

further preferably, the XRPD pattern thereof is substantially in accordance with figure 12;

condition II:

the DSC spectrum shows heat absorption at 191+/-1 ℃;

preferably, the DSC profile thereof substantially corresponds to figure 9; and

condition III:

its TGA profile shows a weight loss of about 0.05% at 179±1 ℃;

preferably, its TGA profile is substantially identical to that of fig. 9.

4. A phosphate of a compound of formula I according to claim 1,

the phosphate of the compound of formula I has form 2 and satisfies at least one of the conditions I to III:

condition I:

its XRPD pattern has characteristic peaks at the following 2θ values: 5.7±0.2°, 11.5±0.2°, 16.8±0.2° and 17.1±0.2°;

preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 3.8±0.2°, 4.9±0.2°, 6.7±0.2°, 11.1±0.2°, 13.6±0.2°, 14.0±0.2°, 19.5±0.2° and 21.3±0.2°;

More preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 9.2±0.2°, 9.8±0.2°, 10.3±0.2°, 12.7±0.2°, 15.7±0.2°, 16.1±0.2° and 22.4±0.2°;

further preferably, the XRPD pattern thereof is substantially in accordance with figure 11;

condition II:

the DSC spectrum shows heat absorption at 119+ -1deg.C, 184+ -1deg.C and 189+ -1deg.C;

preferably, the DSC profile thereof substantially corresponds to figure 8; and

condition III:

its TGA profile shows a weight loss of about 0.79% at 192±1 ℃;

preferably, its TGA profile is substantially identical to that of fig. 8.

5. A phosphate of a compound of formula I according to claim 1,

the phosphate of the compound of formula I has form 3 and satisfies at least one of conditions I-III:

condition I:

its XRPD pattern has characteristic peaks at the following 2θ values: 3.9±0.2°, 4.8±0.2°, 16.5±0.2° and 16.7±0.2°;

preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 7.2±0.2° and 17.5±0.2°;

more preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 11.6+ -0.2 DEG and 14.2+ -0.2 DEG;

further preferably, the XRPD pattern thereof is substantially in accordance with figure 4;

Condition II:

its DSC profile shows endotherms at 183+ -1deg.C and 189+ -1deg.C;

preferably, the DSC profile thereof substantially corresponds to figure 5; and

condition III:

its TGA profile shows a weight loss of about 3.09% at 185±1 ℃;

preferably, its TGA profile is substantially in accordance with figure 5.

6. A phosphate of a compound of formula I according to claim 1,

the phosphate of the compound of formula I has form 4 and satisfies at least one of conditions I-III:

condition I:

its XRPD pattern has characteristic peaks at the following 2θ values: 4.3±0.2°, 4.8±0.2° and 16.5±0.2°;

preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 12.3±0.2°;

more preferably, its XRPD pattern also has characteristic peaks at the following 2θ values: 9.0±0.2°, 9.3±0.2° and 17.6±0.2°;

further preferably, the XRPD pattern thereof is substantially in accordance with figure 10;

condition II:

the DSC spectrum shows heat absorption at 179+ -1deg.C and 187+ -1deg.C;

preferably, the DSC profile thereof substantially corresponds to figure 7; and

condition III:

its TGA profile shows a weight loss of about 1.51% at 189±1 ℃;

preferably, its TGA profile is substantially identical to that of fig. 7.

7. A process for the preparation of a phosphate salt of a compound of formula I according to claim 3, selected from the group consisting of antisolvent processes and suspension-transcrystalline processes, preferably antisolvent processes.

8. The process for the preparation of the phosphate salt of the compound of formula I according to claim 4, which is selected from the group consisting of antisolvent processes and suspension-transcrystalline processes, preferably antisolvent processes.

9. The process for the preparation of the phosphate salt of the compound of formula I according to claim 5, which is a suspension-seeding process.

10. The process for the preparation of the phosphate salt of the compound of formula I according to claim 6, which is selected from the group consisting of antisolvent processes and suspension-transcrystalline processes, preferably antisolvent processes.

11. A pharmaceutical composition comprising a phosphate salt of a compound of formula I according to any one of claims 1 to 6;

12. Use of a phosphate salt of a compound of formula I according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 11 for the manufacture of a medicament for the prevention and/or treatment of intestinal dysfunction caused at least in part by opioid;

preferably, the bowel dysfunction caused at least in part by opioid is constipation.

13. A phosphate salt of a compound of formula I according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 11 for use in the prevention and/or treatment of intestinal dysfunction caused at least in part by opioid;

14. A method for preventing and/or treating bowel dysfunction caused at least in part by opioid comprising the steps of: administering a prophylactically and/or therapeutically effective amount of a phosphate salt of a compound of formula I according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 11 to a subject in need thereof;