WO2023197934A1 - Solid salt form of opioid receptor antagonist conjugate, crystal form, preparation method therefor, composition, and use - Google Patents

Abstract

The present invention relates to the technical field of pharmaceutical crystals, and relates to a solid salt form of an opioid receptor antagonist conjugate, a crystal form, a preparation method therefor, a composition, and a use. Specifically, the conjugate has a structure as shown in formula I, and can form a stable solid phosphate, wherein the molar ratio of phosphoric acid to the compound as shown in formula I is 1:1 to 3:1. The phosphate can be an amorphous substance, or a polymorph having a crystal form 1, 2, 3 or 4. Moreover, the preparation method is simple and feasible, and the present invention is suitable for preventing and treating intestinal dysfunction caused by opioids, such as constipation.

Description

Solid salt forms and crystal forms of opioid receptor antagonist conjugates and preparation methods, compositions and uses thereof

References to related applications

The present invention claims an invention patent submitted in China on April 11, 2022, titled "Solid Salt Form, Crystal Form of Opioid Receptor Antagonist Conjugate and Preparation Method, Composition and Use" and Application Number 202210374703.5 The entire contents of this patent application are incorporated herein by reference.

Technical field

The invention belongs to the technical field of pharmaceutical crystals, and specifically relates to the solid salt form and crystal form of an opioid receptor antagonist conjugate (especially a dual opioid receptor antagonist conjugate), its preparation method, pharmaceutical composition and Medicinal purposes.

Background technique

Cancer pain is one of the most common complications when cancer progresses to the middle and late stages. The incidence of pain in late-stage cancer patients is as high as over 75%. Cancer pain causes huge harm to cancer patients, families and society.

Opioids are the oldest analgesics and the most effective analgesics so far. They have the advantages of strong analgesic effect and no organ toxicity when used for a long time. When a patient's pain worsens due to tumor progression, the analgesic effect can be improved by increasing the drug dose. Therefore, opioid analgesics have an irreplaceable status for patients with moderate and severe cancer pain.

When patients use opioids to treat moderate to severe pain, they may cause many adverse reactions such as nausea, vomiting, and drowsiness. However, the above adverse reactions can usually be tolerated by most patients within a week. However, the incidence rate of constipation caused by opioids is as high as 90% to 100%, and tolerance will not occur due to long-term medication. Constipation not only appears in the initial stage of medication, but also persists throughout the entire process of analgesic treatment. However, it is worth noting that if constipation is not controlled in time, it can cause serious complications and become the biggest obstacle to effective pain relief. At the same time, constipation can seriously affect the treatment of the disease, interrupt the treatment, greatly extend the patient's hospitalization time, and reduce the patient's quality of life. Therefore, preventing and treating constipation adverse reactions is always an issue that cannot be ignored during opioid analgesic treatment.

Research shows that opioid receptors not only exist in the central nervous system (including the brain and spinal cord), but also widely exist in the peripheral nervous system located in organs such as the gastrointestinal tract. Opioids bind to intestinal opioid receptors, causing Intestinal motility is slow, intestinal fluid secretion is reduced, absorption is increased, the activity of excitatory and inhibitory neurons in the intestinal muscularis plexus is reduced, the muscle tension of the intestinal wall smooth muscle is increased and coordinated peristalsis is inhibited, thereby increasing non-peristaltic contractions, and ultimately Cause constipation. Because the body develops tolerance to the intestinal effects of opioids very slowly, constipation will persist during treatment.

Drug therapy is a conventional means to relieve or treat constipation caused by opioids. Specifically, people hope to find an opioid that can block peripheral μ-opioid receptors without affecting the central analgesic effect. Opioid receptor antagonists themselves have no agonistic effect on opiate receptors, but they have strong affinity for mu receptors, and also have certain affinity for kappa, delta and sigma receptors, and can remove binding to these receptors. Binding of opioid analgesics, thereby producing an antagonistic effect. Systemic application of opioid receptor antagonists, such as naloxone, naltrexone, and nalmefene, has effects on both central and peripheral opioid receptors. While antagonizing the peripheral effects of opioids, it also weakens the central analgesic effect.

WO 2017/133634 A1 discloses a series of conjugates of polyethylene glycol and opioid receptor antagonists. Studies have shown that dual opioid receptor antagonist conjugates are more difficult to pass through the blood-brain barrier and can better target the periphery. The nervous system can better antagonize the toxic and side effects of opioids without affecting the analgesic effect of opioids, and has broad clinical application prospects.

In the process of drug formulation and clinical application, it is unanimously expected to use solid drugs with high stability. However, the dual-opioid receptor antagonist conjugate is oily and highly viscous, making it difficult to become a solid, and it is unable to meet the needs for further development of formulations. As for the stable solid salt form and polymorphic form of dual opioid receptor antagonist conjugates, there are no relevant research reports in the prior art.

Contents of the invention

Invent the problem to be solved

In order to ensure the physical and chemical stability of the drug during production, processing and drug storage, through in-depth research on different acid addition salts of dual opioid receptor antagonist conjugates, solid salts and polymorphs that can exist stably are screened out. And the corresponding preparation method is simple, easy and operable.

solutions to problems

In a first aspect, the invention provides phosphate salts of compounds of formula I,

Among them, the molar ratio of phosphoric acid to the compound of formula I is 1:1 to 3:1.

Preferably, in the phosphate salt 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 consistent with Figure 2.

In a second aspect, the present invention provides a phosphate salt of the compound of formula I, which has crystal form 1 and whose 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, 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 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 consistent with Figure 12.

Further, the differential scanning calorimetry (DSC) spectrum of the phosphate of the compound of formula I shows an endotherm at 191±1°C.

Preferably, the DSC spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 9.

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

Preferably, the TGA spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 9.

In a third aspect, the present invention provides a phosphate salt of the compound of formula I, which has crystal form 2 and whose 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, the XRPD pattern of the phosphate 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 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 consistent with Figure 11.

Further, the DSC spectrum of the phosphate of the compound of formula I shows endotherms at 119±1°C, 184±1°C and 189±1°C.

Preferably, the DSC spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 8.

Further, the TGA spectrum of the phosphate salt of the compound of formula I showed a weight loss of about 0.79% at 192±1°C.

Preferably, the TGA spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 8.

In a fourth aspect, the present invention provides a phosphate salt of the compound of formula I, which has crystal form 3 and whose 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, 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° and 17.5±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: 11.6±0.2° and 14.2±0.2°.

Further preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially consistent with Figure 4.

Further, the DSC spectrum of the phosphate of the compound of formula I shows endotherms at 183±1°C and 189±1°C.

Preferably, the DSC spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 5.

Further, the TGA spectrum of the phosphate salt of the compound of formula I shows a weight loss of about 3.09% at 185±1°C.

Preferably, the TGA spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 5.

In a fifth aspect, the present invention provides a phosphate salt of the compound of formula I, which has crystal form 4 and whose XRPD pattern has characteristic peaks at the following 2θ values: 4.3±0.2°, 4.8±0.2° and 16.5±0.2°. .

Preferably, the XRPD pattern of the phosphate salt of the compound of formula I also has a characteristic peak at the following 2θ value: 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 consistent with Figure 10.

Further, the DSC spectrum of the phosphate of the compound of formula I shows endotherms at 179±1°C and 187±1°C.

Preferably, the DSC spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 7.

Further, the TGA spectrum of the phosphate salt of the compound of formula I showed a weight loss of approximately 1.51% at 189±1°C.

Preferably, the TGA spectrum of the phosphate salt of the compound of formula I is substantially consistent with Figure 7.

In a sixth aspect, the present invention provides a method for preparing the phosphate of the compound of formula I described in the second aspect, which is selected from the group consisting of an anti-solvent method and a suspension crystallization method, with the anti-solvent method being preferred.

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 a fatty alcohol, preferably ethanol.

Further, the substance to be crystallized used in the anti-solvent method is the phosphate salt of the compound of formula I described in any one of the first aspect and the third to fifth aspects.

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

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

In the seventh aspect, the present invention provides a method for preparing the phosphate of the compound of formula I described in the third aspect, which is selected from the antisolvent method and the suspension crystallization method, with the antisolvent method being preferred.

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

Further, the substance to be crystallized used in the anti-solvent method is the phosphate salt of the compound of formula I described in any one of the first to second aspects and the fourth to fifth aspects.

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

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

In the eighth aspect, the present invention provides a method for preparing the phosphate of the compound of formula I described in the fourth aspect, which is a suspension crystallization process. Law.

Preferably, the solvent used in the suspension crystallization method is fatty ether, fatty ketone, fatty acid ester, 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 salt of the compound of formula I described in the first aspect.

In the ninth aspect, the present invention provides a method for preparing the phosphate of the compound of formula I described in the fifth aspect, which is selected from the antisolvent method and the suspension crystallization method, with the antisolvent method being preferred.

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 fatty acid ester, preferably methyl formate.

Further, the substance to be crystallized used in the anti-solvent method is the phosphate salt of the compound of formula I described in any one of the first to fourth aspects.

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

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

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

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

In the eleventh aspect, the present invention provides the phosphate salt of the compound of formula I according to any one of the first to fifth aspects or the pharmaceutical composition in the tenth aspect when prepared for the prevention and/or treatment of, at least in part, Use in medicines for intestinal disorders caused by opioids.

In the twelfth aspect, the present invention provides the phosphate salt of the compound of formula I according to any one of the first to fifth aspects or the pharmaceutical composition in the tenth aspect, which is used for preventing and/or treating at least part of the disease caused by Intestinal disorders caused by opioids.

In a thirteenth aspect, the present invention provides a method for preventing and/or treating intestinal dysfunction caused at least in part by opioids, comprising the steps of: adding a preventive and/or therapeutically effective amount of the first aspect to The phosphate salt of the compound of formula I according to any one of the fifth aspects or the pharmaceutical composition according to the tenth aspect is administered to an individual in need thereof.

Further, the intestinal dysfunction caused at least in part by opioids is constipation.

Effect of invention

By screening acid addition salts of opioid receptor antagonist conjugates, the present invention obtains solid bisphosphates and their polymorphs that can exist stably. Among them, crystal form 1 has excellent thermodynamic stability.

Description of the drawings

Figure 1 is the ¹ H-NMR spectrum of PEG8-(6-α-naltrexide)2 free base.

Figure 2 is the XRPD pattern of the amorphous substance in Example 2.

Figure 3 is a PLM photograph of the amorphous material in Example 2.

Figure 4 is the XRPD pattern of the polymorph (form 3) prepared in the methyl tert-butyl ether-methyl formate system in Example 2.

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

Figure 6 is the XRPD overlay pattern of the two polymorphs obtained under different acid-base molar ratios in Example 2.

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

Figure 8 is the DSC and TGA overlay spectrum of the polymorph (form 2) prepared in the methanol-acetone system in Example 2.

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

Figure 10 is the XRPD pattern of the polymorph (form 4) prepared in the methanol-methyl formate system in Example 2.

Figure 11 is the XRPD pattern of the polymorph (form 2) prepared in the methanol-ethanol-tetrahydrofuran system in Example 2.

Figure 12 is the XRPD pattern of the polymorph (form 1) prepared in the ethanol-water system in Example 2.

Figure 13 is the XRPD overlay pattern of the polymorphs prepared in different systems in Example 2.

Figure 14 is an XRPD overlay pattern showing the preliminary research results of suspension crystallization in Example 2.

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

Figure 16 is the XRPD overlay pattern of the four polymorphs produced by the optimized process in Example 2.

Figure 17 is an XRPD overlay pattern showing the crystallization state of the polymorph (form 2) in a water-ethanol system (conversion to crystal form 1).

Figure 18 is an XRPD overlay pattern showing the crystallization state of the polymorph (form 3) in the water-ethanol system (transformation from the intermediate state of form 2 to form 1).

Figure 19 is a DSC overlay pattern showing the crystallization state of the polymorph (form 3) in a water-ethanol system (transformation from the intermediate state of form 2 to form 1).

Figure 20 is an XRPD overlay pattern showing the crystallization state of the polymorph (form 4) in a water-ethanol system (conversion to crystal form 1).

Figure 21 is an XRPD overlay pattern showing the crystallization state of the polymorph (form 1) in the methanol-acetone system (no crystallization occurs).

Figure 22 is an XRPD overlay pattern showing the crystal transformation of the polymorph (form 1) under high temperature and mechanical grinding conditions (no crystal transformation occurred).

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

Detailed ways

Unless otherwise defined, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art.

Unless otherwise stated, words in the singular such as "a," "an," and "the" appearing herein encompass their plural counterparts unless the context clearly dictates otherwise. For example, when referring to "a" crystal or polymorph, this encompasses one or more different crystals or polymorphs, and when referring to "the" method, this encompasses ordinary skill in the art. Equivalent procedures and methods known to persons.

Unless otherwise stated, the term “comprises” and variations thereof, such as “contains” and “includes”, appear herein to mean that the collection not only encompasses the expressly disclosed one or more integers, steps, or combinations thereof, but also excludes any Other integers, steps, or combinations thereof. At the same time, under certain circumstances, the term "comprising" appearing in this article may be replaced by the term "contains," "includes," or "having."

For the crystalline forms in this article, only the characteristic peaks in the XRPD pattern (i.e., the most characteristic, significant, unique and/or repeatable diffraction peaks) are summarized, while other peaks can be obtained from the pattern by conventional methods. obtained in. The above characteristic peaks can be repeated within the error limit (the error range is ±0.2°).

Solid salts of opioid receptor antagonist conjugates

In view of the problem that opiate receptor antagonist conjugates are highly viscous, difficult to solidify, and cannot meet formulation requirements, the present invention hopes to obtain a solid suitable for formulations by forming an acid addition salt with an acid. After a large number of salt form and crystal form screening experiments, the present invention successfully discovered phosphates (especially diphosphates) that can stably exist in solid form (especially crystal form).

The present invention provides solid salts of opioid receptor antagonist conjugates, in which the free base part can be PEG8-(6-α-naltrexine)2 (having the structure shown in formula I), and the acid part can be phosphoric acid .

In one embodiment, the solid salt of the above-mentioned opioid receptor antagonist conjugate is a phosphate salt of PEG8-(6-α-naltrexine)2, wherein the molar ratio of phosphoric acid to free base can be 1:1~ 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 within the above range.

In one embodiment, the molar ratio of phosphoric acid to free base in the solid salt of the above-mentioned opioid receptor antagonist conjugate can be 1.5:1 to 2.5:1, such as 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 range.

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

In one embodiment, the solid salt of the above-mentioned opioid receptor antagonist conjugate is the phosphate salt of PEG8-(6-α-naltrexine)2, which may exist in an amorphous form. Under certain conditions, the XRPD pattern of the amorphous substance can be basically consistent with Figure 2.

In another embodiment, the solid salt of the above-described opioid receptor antagonist conjugate is the phosphate salt of PEG8-(6-alpha-naltrexine)2, which may exist in the form of polymorphs. The polymorph can have multiple crystal forms and can be characterized by at least one of XRPD, DSC and TGA.

In one embodiment, the polymorph described above may have Form 1. The XRPD pattern of the polymorph having Form 1 may 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°, and preferably may also have the following Characteristic peaks at 2θ values: 13.2±0.2°, 13.6±0.2°, 16.7±0.2°, 21.3±0.2° and 22.7±0.2°, and more preferably, 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 further preferably can be basically consistent with Figure 12.

And/or, the DSC pattern of the polymorph having Form 1 may show an endotherm at 191 ± 1° C., and preferably may be substantially consistent with FIG. 9 .

And/or, the TGA pattern of the polymorph having Form 1 may show a weight loss of about 0.05% at 179 ± 1° C., and preferably may be substantially consistent with FIG. 9 .

In another embodiment, the polymorph described above may have Form 2. The XRPD pattern of the polymorph with Form 2 may 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°, and preferably may also have the following Characteristic peaks at 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 It may 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 it may be substantially The above is consistent with Figure 11.

And/or, the DSC pattern of the polymorph having Form 2 may show endotherms at 119±1°C, 184±1°C and 189±1°C, and preferably may be substantially consistent with FIG. 8 .

And/or, the TGA pattern of the polymorph having Form 2 may show a weight loss of about 0.79% at 192±1°C, and preferably may be substantially consistent with FIG. 8 .

In yet another embodiment, the polymorph described above may have Form 3. The XRPD pattern of the polymorph with crystalline form 3 may 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°, and preferably may also have the following Characteristic peaks at 2θ values: 7.2±0.2° and 17.5±0.2°. More preferably, it can also have characteristic peaks at the following 2θ values: 11.6±0.2° and 14.2±0.2°. Further preferably, it can be basically consistent with Figure 4 .

And/or, the DSC pattern of the polymorph having Form 3 may show endotherms at 183±1°C and 189±1°C, and preferably may be substantially consistent with Figure 5 .

And/or, the TGA pattern of the polymorph having Form 3 can show a weight loss of about 3.09% at 185±1°C, and preferably can be substantially consistent with Figure 5 .

In yet another embodiment, the polymorph described above may have Form 4. The XRPD pattern of the polymorph with Form 4 may have characteristic peaks at the following 2θ values: 4.3±0.2°, 4.8±0.2° and 16.5±0.2°, and preferably may also have characteristics at the following 2θ values. Peak: 12.3±0.2°. More preferably, it can also have characteristic peaks at the following 2θ values: 9.0±0.2°, 9.3±0.2° and 17.6±0.2°. Further preferably, it can be basically consistent with Figure 10.

And/or, the DSC pattern of the polymorph having Form 4 may show endotherms at 179±1°C and 187±1°C, and preferably may be substantially consistent with Figure 7 .

And/or, the TGA pattern of the polymorph having Form 4 can show a weight loss of about 1.51% at 189±1°C, and preferably can be substantially consistent with Figure 7 .

Methods for Preparing Solid Salts of Opioid Receptor Antagonist Conjugates

The present invention also provides various preparation methods of solid salts of the above-mentioned opioid receptor antagonist conjugates (eg, bisphosphate salts of PEG8-(6-α-naltrexine)2).

In one embodiment, the above-mentioned diphosphate of PEG8-(6-α-naltrexine)2 can be prepared by the following method: respectively dissolving phosphoric acid and PEG8-(6-α-naltrexine)2 in a solvent , to obtain an acid solution and a free base solution; then mix the acid solution and the free base according to the proportion and stir, and obtain the diphosphate of PEG8-(6-α-naltrexine) 2 after desolvation.

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

The present invention does not impose any clear limitation on the concentration of the acid solution and/or the free base solution in the above method. For example, the concentration of the acid solution may be 0.01 to 1 mol/L (especially 0.1 mol/L).

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

The present invention does not impose any clear restrictions on the stirring conditions (for example, temperature, duration, etc.) in the above method. For example, the stirring conditions can be stirred at room temperature for 15 minutes.

The present invention does not impose any clear limitations on the desolvation conditions (for example, temperature, pressure, duration, etc.) in the above method. For example, it can be carried out by distillation under reduced pressure (for example, using a rotary evaporator).

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

In the present invention, the diphosphate of PEG8-(6-α-naltrexane)2 prepared by the above method can exist in the form of an amorphous substance, and its specific characterization method and results are as described above.

In addition, the present invention also provides a method for preparing the diphosphate of PEG8-(6-α-naltrexine) 2 in the form of polymorphs.

First, the polymorph of the above-mentioned PEG8-(6-α-naltrexine) 2 bisphosphate has crystal form 1, and its preparation method can be an anti-solvent method or a suspension crystallization method, and preferably it can be an anti-solvent method. .

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

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

In one embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexide)2 in the form of an amorphous substance.

In another embodiment, the substance to be crystallized used in the antisolvent method described above may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 2.

In yet another embodiment, the substance to be crystallized used in the above-mentioned antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 3.

In yet another embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 4.

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

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

In another embodiment, the substance to be crystallized used in the above-mentioned suspension crystallization method can be the diphosphate salt of PEG8-(6-α-naltrexine) 2 existing in the form of a polymorph with crystal form 2 .

In yet another embodiment, the substance to be crystallized used in the above-mentioned suspension crystallization method may be a diphosphate salt of PEG8-(6-α-naltrexine) 2 existing in the form of a polymorph with crystal form 3. .

In yet another embodiment, the substance to be crystallized used in the above-mentioned suspension crystallization method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 existing in the form of a polymorph with crystalline form 4. .

Secondly, the polymorph of the above-mentioned PEG8-(6-α-naltrexine) 2 bisphosphate has crystal form 2, and its preparation method can be an anti-solvent method or a suspension crystallization method, and preferably it can be an anti-solvent method. .

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

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

In one embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexide)2 in the form of an amorphous substance.

In another embodiment, the substance to be crystallized used in the antisolvent method described above may be the diphosphate salt of PEG8-(6-α-naltrexine)2 in the form of a polymorph having Form 1.

In yet another embodiment, the substance to be crystallized used in the above-mentioned antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 3.

In yet another embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 4.

In one embodiment, the solvent used in the above-mentioned suspension crystallization method can be water, fatty alcohol, fatty ketone or any combination thereof, preferably an aqueous ethanol solution or acetone with a volume ratio of alcohol to water of 20:1.

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

In another embodiment, the substance to be crystallized used in the above-mentioned suspension crystallization method can be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph with crystal form 3. .

Thirdly, the polymorph of the above-mentioned PEG8-(6-α-naltrexine) 2 bisphosphate has crystal form 3, and its preparation method can be a suspension crystallization method.

In one embodiment, the solvent used in the above-mentioned suspension crystallization method can be fatty ether, fatty ketone, fatty acid ester, fatty nitrile or any combination thereof, preferably it can be methyl tert-butyl ether-methyl formate, acetone or Acetonitrile, more preferably, can be methyl tert-butyl ether-methyl formate.

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

Fourth, the polymorph of the above-mentioned PEG8-(6-α-naltrexine) 2 bisphosphate has crystal form 4, and its preparation method can be an anti-solvent method or a suspension crystallization method, and preferably it can be an anti-solvent method. .

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

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

In one embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexide)2 in the form of an amorphous substance.

In another embodiment, the substance to be crystallized used in the antisolvent method described above may be the diphosphate salt of PEG8-(6-α-naltrexine)2 in the form of a polymorph having Form 1.

In yet another embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 2.

In yet another embodiment, the substance to be crystallized used in the above antisolvent method may be the diphosphate salt of PEG8-(6-α-naltrexine) 2 in the form of a polymorph having Form 3.

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

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

Pharmaceutical compositions containing solid salts of opioid receptor antagonist conjugates

In the present invention, the solid salt of the above-mentioned opioid receptor antagonist conjugate can be used alone or in combination with other substances. Therefore, the present invention also provides a pharmaceutical composition, which can contain any of the substances of the present invention. Solid salts of opioid receptor antagonist conjugates existing in an amorphous form (especially in the form of an amorphous substance or a polymorph having any one of the crystalline forms 1 to 4).

In one embodiment, the above pharmaceutical composition may further comprise at least one pharmaceutically acceptable auxiliary material, which auxiliary material has a well-known connotation definition in the art.

Pharmaceutical uses of solid salts of opioid receptor antagonist conjugates or pharmaceutical compositions containing the same

In the present invention, whether it is a solid salt of the opioid receptor antagonist conjugate in any form (especially in the form of an amorphous substance or a polymorph having any one of crystal forms 1 to 4), Either the pharmaceutical composition containing the solid salt can be used to prevent and/or treat intestinal dysfunction caused at least in part by opioids.

Thus, in one aspect, the present invention provides solids of opioid receptor antagonist conjugates in any form, in particular in the form of an amorphous form or a polymorph having any one of Forms 1 to 4. Use of a salt or a pharmaceutical composition containing the same for the preparation of a medicament for preventing and/or treating intestinal dysfunction caused at least in part by opioids. On the other hand, the present invention also provides a method for preventing and/or treating intestinal dysfunction caused at least in part by opioids, which may include the following steps: administering a preventive and/or therapeutic effective amount in any form A solid salt of an opioid receptor antagonist conjugate (especially in the form of an amorphous substance or a polymorph having any one of crystal forms 1 to 4) or a pharmaceutical composition containing the same is administered to the Individuals in need.

In one embodiment, the above-mentioned intestinal disorder caused at least in part by opioids can be constipation.

The present invention will be further illustrated below through specific examples. Unless otherwise stated, the instruments, materials, materials, etc. used in the following examples can be obtained through conventional commercial means.

Analytical methods and instruments

(1) Hydrogen nuclear magnetic resonance spectrum ( ¹ H-NMR)

¹ H-NMR spectra were collected using a Bruker Advance 300M nuclear magnetic resonance instrument equipped with a B-ACS 120 automatic sampling system.

(2)X-ray powder diffraction (XRPD)

X-ray powder diffraction uses a Bruker D8 advance powder X-ray diffractometer and a Bruker D2 phaser powder X-ray diffractometer equipped with a LynxEye detector. The Bruker D8 advance powder X-ray diffractometer was used to test the 2θ scanning angle of the sample from 3° to 40°, and the scanning step was 0.02°. The light tube voltage and light tube current when measuring the sample were 40kV and 40mA respectively. The Bruker D2 phaser powder X-ray diffractometer was used to test the 2θ scanning angle of the sample from 3° to 40°, and the scanning step was 0.02°. The light tube voltage and light tube current when measuring the sample were 30kV and 10mA respectively.

(3)Polarized light microscopy analysis (PLM)

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

(4) Thermogravimetric analysis (TGA)

Thermogravimetric analysis uses TGA Q500 thermogravimetric analyzer or Discovery TGA 55 thermogravimetric analyzer. The test sample is placed in a balanced open aluminum sample pan, and the mass is automatically weighed in the TGA heating furnace. The sample was heated to the final temperature at a heating rate of 10°C/min.

(5) Differential scanning calorimetry (DSC)

Differential scanning calorimetry analysis uses DSC Q200 differential scanning calorimeter or Discovery DSC 250 differential scanning calorimeter. After being accurately weighed, the test sample is placed in the DSC punched sample pan and the exact mass of the sample is recorded. The sample was heated to the final temperature at a heating rate of 10°C/min.

(6) Ion chromatography analysis (IC)

Ion chromatography analysis used a Thermo Fischer ICS-5000+ ion chromatography system, the chromatographic column was IonPac ^TM AS11 (4*250mm), the mobile phase was 30mM KOH aqueous solution, and the flow rate was 1.0mL/min. Accurately weigh 25±2mg of the sample into a 100mL volumetric flask, add 2/3 volume of water to dissolve (ultrasonic if necessary), then dilute to volume with water and mix evenly to obtain a sample solution. Use the mobile phase to balance the chromatography system and the chromatographic column to the baseline balance. According to the blank solution (water), the quantitative limit solution (based on phosphate ions, the concentration is 4 μg/mL), and the working standard solution (based on phosphate ions, the concentration is 40 μg /mL, repeat the injection 6 times), retest standard solution (concentration is the same as the working standard solution), blank solution, sample solution, and working standard solution are injected sequentially, and the content of phosphate ions in the sample is calculated according to the following formula:

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

A(std): The average value of the phosphate ion peak area in the initial six continuous working standard solutions;

Conc.(std): Concentration of phosphate ions in the phosphate ion standard solution (1000μg/mL);

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

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

DF(std): Dilution factor (25) of the working standard.

Solvents used and their abbreviations

Methanol (MeOH), toluene, methyl tert-butyl ether (MTBE), ethyl ketone (MEK), methyl formate (MF), acetone, ethanol (EtOH), acetonitrile (ACN), ethyl acetate (EA), diacetate Methyl chloride (DCM), tetrahydrofuran (THF), isopropyl alcohol (IPA), water (W).

Example 1: Screening of Salt Forms of Opioid Receptor Antagonist Conjugates

1.96-well plate salt screening

14 kinds of 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 Dissolve in methanol and prepare a 0.1mol/L acid solution.

Take another PEG8-(6-α-naltrexine)2 free base (prepared according to the method described in WO 2017/133634 A1, and its ¹ H-NMR spectrum is shown in Figure 1), dissolve it in methanol, and prepare 30 mg /mL solution, use this solution to plate on a 96-well plate, spread three plates in parallel, and add 100 μL to each well.

Using hydrochloric acid, hydrobromic acid, sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, maleic acid and phosphoric acid as acid A, set up two groups in parallel. The first group added 30 μL/well to the first 96-well plate. acid solution so that the molar ratio of acid A to free base is about 1:1. The second group added 60 μL/well of the acid solution to the second 96-well plate so that the molar ratio of acid A to free base is about 2: 1. Using boric acid, palmitic acid, fumaric acid, citric acid, succinic acid and L-tartaric acid as acid B, add 60 μL/well of acid solution to the third 96-well plate so that the molar ratio of acid B to free base is approximately 2:1.

After the methanol has completely evaporated, add 200 μL of solvent to each well, seal it with a perforated sealing film, and place it in a fume hood at room temperature to allow the solvent to evaporate. The specific results are shown in Table 1.

Table 1. Screening results of salt formation of opioid receptor antagonist conjugate free base and various acids in various solvents

The results showed that after the solvent in the 96-well plate evaporated, most of the samples in the wells were glassy oil. Among them, phosphate obtained a solid in MF and THF; palmitate obtained a mixture of oil and solid in MTBE, MF, and EtOH; L-tartrate obtained a solid in acetone and ACN, but after repeated All samples were oily during the experiment, and the process reproducibility was low; the remaining samples were all oily.

2. Study on the salt formation of phosphoric acid and free base under different ratios

Add the methanol solution containing 0.1 mol/L phosphoric acid to the methanol solution containing free base (the molar ratio of acid to alkali is 1:1 and 2:1 respectively), stir at room temperature for 15 minutes and then spin dry to obtain an off-white solid. Add a mixture solution of ethanol and water (the ratio of alcohol to water is about 150:1), raise the temperature to 45°C to dissolve the solution, then naturally cool to room temperature, stir overnight, and a white solid will precipitate. XRPD was used for characterization (as shown in Figure 6). The results showed that the same crystal (form 1) was obtained under two acid-base ratios; through ion chromatography analysis, the content of phosphate ions in both crystals was approximately 15.6 wt%, indicating that the free base and phosphoric acid form an acid addition salt at a molar ratio of 1:2.

Example 2: Screening of crystal forms of opioid receptor antagonist conjugate phosphates

1. Preparation and characterization of starting materials

In this example, two forms of phosphate, crystal and amorphous, were prepared as starting materials for crystal form screening.

Phosphate amorphous substance: first react with PEG8-(6-α-naltrexine)2 free base (prepared according to the method described in WO 2017/133634 A1) and phosphoric acid in methanol (molar ratio of acid to free base Approximately 1:1), a methanol solution of phosphate was obtained, and then the solution was evaporated to dryness by a rotary evaporator, and characterized by XRPD (as shown in Figure 2) and PLM (as shown in Figure 3), which is not seen in the spectrum. There are obvious diffraction peaks, and there are no obvious crystal particles in the photo, which is consistent with the characteristics of amorphous matter.

Phosphate polymorph: Suspend the amorphous phosphate prepared above in the MTBE-MF system and stir overnight to obtain the phosphate polymorph (form 3), which is characterized by XRPD, DSC and TGA. The results are shown in Table 2 and Figure 4-5.

Table 2. XRPD characterization results of Form 3

2. Preliminary crystal form screening

For the phosphate amorphous substance or polymorph obtained in item 1, parallel experiments were conducted using MTBE, MeOH and water as good solvents, and MF, THF and EtOH as anti-solvents, and four different crystals were obtained. Salt (phosphate polymorph), the specific preparation method is shown in Table 3.

Table 3. Preparation method of opiate receptor antagonist conjugate crystal salts

As can be seen from Figure 9 (Crystal Form 1), the TGA spectrum of the crystalline salt obtained in the ethanol-water system shows a small weight loss (0.05%) before decomposition, and the DSC spectrum shows that the endothermic peak is a single peak. , the results are better than the crystalline salts obtained in the other two systems. It can be seen from Figure 7 (Crystal Form 4) and Figure 23 (Crystal Form 2) that the TGA spectra of the crystalline salts obtained in the other two systems show more weight loss, and the DSC spectra show that the endothermic peaks are not single. Peaks indicate that crystal transfer or mixed crystals may occur during the heating process. For products obtained using other good solvent-antisolvent systems, their crystallinity is very poor, and they are basically mixtures of amorphous substances and crystals or oily substances, which will not be described in detail here.

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

Table 4. XRPD characterization results of Form 4

Table 5. XRPD characterization results of Form 2

Table 6. XRPD characterization results of Form 1

From the above results and Figure 13, it can be seen that the four types of crystal salts (phosphate polymorphs) obtained by the above method have different crystal forms.

3. Preliminary study on suspension crystallization (transformation of amorphous matter into polymorphic form)

Use the phosphate amorphous material prepared in item 1 to conduct a single solvent suspension crystallization study. The specific method is as follows: weigh about 200mg of the sample, add 3mL of solvent, stir at room temperature for 2 days, and perform XRPD analysis after sampling. The results are as shown in the figure 14 shown. The results show that after suspension crystallization in ethanol, ethyl acetate, methylene chloride, methyl ethyl ketone, isopropyl alcohol, methyl tert-butyl ether and toluene, the phosphate remains amorphous or approximates an amorphous substance. Product; after suspension crystallization in tetrahydrofuran and methyl formate, the same new phosphate polymorph (form 4) was obtained; after suspension crystallization in acetone, another new polymorph was obtained Form (form 2), but its crystallinity is low; after suspension crystallization in acetonitrile, the phosphate polymorph (form 3) prepared in item 1 is obtained, but its crystallinity is too low.

4. Research on optimization of crystallization process

Since the structure of PEG8-(6-α-naltrexine)2 free base contains a flexible PEG chain, its phosphate is difficult to crystallize. Under most conditions in item 3, only amorphous or amorphous products can be obtained. Low crystallinity polymorph. Therefore, based on the aforementioned research results, the phosphate amorphous substance or crystal prepared in item 1 was used as the starting material, an appropriate good solvent-antisolvent system was selected to optimize the crystallization process, and methanol and water were finally selected as Good solvents include methyl formate, ethanol, tetrahydrofuran, acetone, ethyl acetate and dichloromethane as anti-solvents. The details are shown in Table 7.

Table 7. Optimized preparation process of opiate receptor antagonist conjugate crystal salts

It can be seen from Figure 15 that the polymorph with crystal form 1 can be obtained in the ethanol-water system and has good crystallinity; it can be obtained in both the methyl formate-water system and the methyl formate-methanol system. Polymorphs of crystal form 4, but the crystallinity is relatively low; polymorphs with crystal form 2 can be obtained in the methanol-acetone system, methanol-ethyl acetate system and methanol-ethanol-tetrahydrofuran system, and It has good crystallinity; in the methanol-dichloromethane system, only amorphous or nearly amorphous products can be obtained.

The optimized process for the preparation of 4 phosphate polymorphs with different crystal forms was finally determined:

Weigh a certain amount of phosphate sample (for example, 61.2 mg of the phosphate polymorph (crystalline form 3) prepared in item 1), and add water and ethanol (for example, 0.06 mL of water, 1 mL of ethanol) successively. ), an oily substance begins to appear, and then it is heated to 40°C and stirred (for example, 3 hours), then slowly lowered to room temperature and stirred (for example, 24 hours), and filtered with suction to obtain white crystals (crystalline form 1).

Weigh a certain amount of phosphate sample (for example, 197.6mg of the phosphate amorphous substance prepared in item 1), add methanol (for example, 0.4mL) to dissolve, and then slowly add acetone (for example, 4mL) to precipitate the solid , stir at room temperature (for example, overnight), and filter with suction to obtain white crystals (crystal form 2).

Weigh a certain amount of phosphate sample (for example, phosphate amorphous material prepared by the following method: weigh 8.8g of free base methanol solution (14.47wt%), add 14mL of 0.1mol/L phosphoric acid methanol solution, and react for 15 minutes Then, spin the solvent dry to obtain), add a mixed solution of methyl formate and methyl tert-butyl ether (for example, 2 mL of methyl formate, 10 mL of methyl tert-butyl ether), and stir at room temperature (for example, overnight) , suction filtration, and white crystals (form 3) were obtained.

Weigh a certain amount of phosphate sample (for example, phosphate amorphous material prepared by the following method: weigh 3.5g of free base methanol solution (14.47wt%), add 5.2mL of 0.1mol/L phosphoric acid methanol solution, react After 30 minutes, spin dry the solvent. to obtain), add methanol (for example, 0.5mL), after the oily substance appears, add methyl formate (for example, 7.5mL), the oily substance turns into a solid, then add methyl formate (for example, 5mL), and stir at room temperature ( For example, 1 hour), filter with suction, and obtain white crystals (crystal form 4).

XRPD analysis was performed on the above four polymorphs having crystal forms 1-4 respectively, and the results are shown in Figure 16. From the XRPD pattern, crystal forms 3 and 4 show lower crystallinity, and crystal forms 1 and 2 show better crystallinity.

5. In-depth research on suspension crystallization (transformation between different polymorphs)

(1) Crystal transformation of Form 2 in ethanol-water system

The phosphate polymorph (crystal form 2) sample was added to an ethanol system with a water content of 1% and stirred at 40°C for 2 days to obtain a solid sample. XRPD detection showed that it had completely transformed into crystal form 1 (as shown in Figure 17 Show).

(2) Crystal transformation of Form 3 in ethanol-water system

Take approximately 300 mg of the phosphate polymorph (crystal form 3), add a mixed solution of ethanol and water (containing 0.15 ml of water and 3 ml of ethanol), and stir at room temperature for 4 hours. XRPD detection shows a change in the crystal form; raise the temperature to 40°C. By 8 hours, XRPD detection showed that it had completely transformed into crystal form 2; by 12 hours, crystal form 2 began to transform into crystal form 1; by 30 hours, XRPD detection showed that it had completely transformed into crystal form 1 (such as As shown in Figure 18).

The DSC results also reflect the transformation process from crystal form 3 to crystal form 1, which shows a gradual transformation from two endothermic peaks to one endothermic peak (as shown in Figure 19).

(3) Crystal transformation of Form 4 in ethanol-water system

The phosphate polymorph (crystal form 4) sample was added to an ethanol system with a water content of 1% and stirred at 40°C for 2 days to obtain a solid sample. XRPD detection showed that it had completely transformed into crystal form 1 (as shown in Figure 20 Show).

(4) Study on crystallization of Form 1 in methanol-acetone system

The aforementioned research results show that crystal forms 2-4 can be transformed into crystal form 1 under certain conditions, which initially indicates that crystal form 1 may be the dominant crystal form. Since the specific type of solvent may affect the transformation of the crystal form, suspension crystallization was used to further investigate whether crystallization of Form 1 occurred in other systems.

The phosphate polymorph (crystal form 1) sample was added to the methanol-acetone system for suspension and crystallization, and was beaten and stirred for 2 days to obtain a solid sample. XRPD detection showed no change (as shown in Figure 21). The results show that even if crystal form 1 is suspended and crystallized under the process conditions for preparing crystal form 2, it cannot be transformed into crystal form 2. The stability of crystal form 1 is relatively high.

6. Study on the stability of crystal form 1 under high temperature and mechanical grinding conditions

High-temperature stability study: Bake the phosphate polymorph (form 1) sample in an oven at 65°C for 3 days, and perform XRPD detection after sampling.

Mechanical grinding stability: Mechanically grind the phosphate polymorph (crystal form 1) sample for 3 minutes, and perform XRPD detection after sampling.

XRPD detection also showed no change (as shown in Figure 22), further indicating that crystal form 1 has high stability and is suitable for subsequent formulation development.

Claims (12)

1. Phosphate salts of compounds of formula I,
   

   Among them, the molar ratio of phosphoric acid to the compound of formula I is 1:1 to 3:1, preferably 1.5:1 to 2.5:1, and more preferably 2:1.
2. The phosphate salt of the compound of formula I according to claim 1, characterized in that,

   The phosphate salt of the compound of formula I is amorphous;

   Preferably, the XRPD pattern of the phosphate salt of the compound of formula I is substantially consistent with Figure 2.
3. The phosphate salt of the compound of formula I according to claim 1, characterized in that,

   The phosphate salt of the compound of formula I has crystal form 1 and meets at least one of 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, its XRPD pattern is basically consistent with Figure 12;

   Condition II:

   Its DSC spectrum shows endotherm at 191±1°C;

   Preferably, its DSC pattern is substantially consistent with Figure 9; and

   Condition III:

   Its TGA pattern shows about 0.05% weight loss at 179±1°C;

   Preferably, its TGA pattern is substantially consistent with Figure 9.
4. The phosphate salt of the compound of formula I according to claim 1, characterized in that,

   The phosphate salt of the compound of formula I has crystal form 2 and meets at least one of 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, its XRPD pattern is basically consistent with Figure 11;

   Condition II:

   Its DSC pattern shows endotherms at 119±1°C, 184±1°C and 189±1°C;

   Preferably, its DSC pattern is substantially consistent with Figure 8; and

   Condition III:

   Its TGA pattern shows about 0.79% weight loss at 192±1°C;

   Preferably, its TGA pattern is substantially consistent with Figure 8.
5. The phosphate salt of the compound of formula I according to claim 1, characterized in that,

   The phosphate salt of the compound of formula I has crystal form 3 and meets at least one of conditions I to 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° and 14.2±0.2°;

   Further preferably, its XRPD pattern is basically consistent with Figure 4;

   Condition II:

   Its DSC spectrum shows endotherms at 183±1°C and 189±1°C;

   Preferably, its DSC pattern is substantially consistent with Figure 5; and

   Condition III:

   Its TGA pattern shows about 3.09% weight loss at 185±1°C;

   Preferably, its TGA pattern is substantially consistent with Figure 5.
6. The phosphate salt of the compound of formula I according to claim 1, characterized in that,

   The phosphate salt of the compound of formula I has crystal form 4 and meets at least one of conditions I to 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, its XRPD pattern is basically consistent with Figure 10;

   Condition II:

   Its DSC spectrum shows endotherms at 179±1°C and 187±1°C;

   Preferably, its DSC pattern is substantially consistent with Figure 7; and

   Condition III:

   Its TGA pattern shows about 1.51% weight loss at 189±1°C;

   Preferably, its TGA pattern is substantially consistent with Figure 7.
7. The preparation method of the phosphate of the compound of formula I according to any one of claims 3, 4 and 6, which is selected from the anti-solvent method and the suspension crystallization method, with the anti-solvent method being preferred.
8. The preparation method of the phosphate of the compound of formula I according to claim 5, which is a suspension crystallization method.
9. A pharmaceutical composition comprising a phosphate salt of the compound of formula I according to any one of claims 1 to 6;

   Preferably, the pharmaceutical composition further contains at least one pharmaceutically acceptable excipient.
10. The phosphate salt of the compound of formula I according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 9 for use in the prevention and/or treatment of intestinal dysfunction caused at least in part by opioids Use in medicines;

    Preferably, the intestinal disorder caused at least in part by opioids is constipation.
11. The phosphate salt of the compound of formula I according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 9 for preventing and/or treating intestinal dysfunction caused at least in part by opioids ;

    Preferably, the intestinal disorder caused at least in part by opioids is constipation.
12. A method for preventing and/or treating intestinal dysfunction caused at least in part by opioids, comprising the steps of: adding a preventive and/or therapeutically effective amount of a drug according to any one of claims 1 to 6 The phosphate salt of the compound of formula I or the pharmaceutical composition according to claim 9 is administered to an individual in need thereof;

    Preferably, the intestinal disorder caused at least in part by opioids is constipation.