Metal complexes of FMOC-protected lipoPEG-peptides and metal halide salts and preparation thereof

Abstract

The present invention relates to the general field of amino acids and peptides. In particular, the present invention relates to solid metal complex salts of amino acids or peptides, to processes for preparing such solid metal complex salts and to the use of such salts in the preparation of semeglutide, tiall and pharmaceutically acceptable salts thereof.

Description

TECHNICAL FIELD

[0001] The present invention relates to the general field of amino acids and peptides. In particular, the present invention relates to solid metal complex salts of amino acids or peptides, to a process for manufacturing such solid metal complex salts, and to the use of the salts in the preparation of semaglutide, tirzepatide and pharmaceutically acceptable salts thereof. BACKGROUND

[0002] Amino acids and peptides that contain both a hydrophilic moiety and an alkylated moiety in the side chain typically present themselves as viscous oils, gels or waxes; this physical state makes them very difficult to handle. Simple operations, such as weighing or transferring them from one container or vessel to another, for purification, characterization or further reaction, all result in substantial loss of material.

[0003] For example, the compounds of Formula 1, Formula 2, Formula 3 and Formula 4 are in this situation: Formula 1 Formula 2 Formula 3 Formula 4.

[0004] The compounds of Formula 1 and Formula 3 are intermediates in the synthesis of semaglutide, a human GLP-1 receptor agonist indicated for the adjunct to diet and exercise to improve glycemic control in adult patients with type 2 diabetes mellitus, as well as for the chronic management of body weight in overweight or obese adult patients.

[0005] The compounds of Formula 2 and Formula 4 are intermediates in the synthesis of tirzepatide, which is used in the same therapeutic area as semaglutide. The compound of Formula 2 differs from the compound of Formula 1 in the presence of a C20 aliphatic hydrocarbon chain instead of a C18 aliphatic hydrocarbon chain.

[0006] The use of compounds analogous to Formula 1-Formula 4 for the preparation of semaglutide and tirzepatide is well known in the art.

[0007] WO2019170895A1 describes the synthesis of GLP-1 compounds by applying an enzymatic approach, in which the target product is generated by enzyme catalyzed condensation of two corresponding fragments. The process to generate these fragments requires the use of, for example, a compound of Formula 1 to generate one of the fragments used in the enzymatic synthesis of said semaglutide.

[0008] WO2017114191A1 provides another example in which a compound of formula 3 was prepared and used as an intermediate in the synthesis of smegglutinin.

[0009] However, the compounds of formulas 1-4 are themselves in the form of pale yellow viscous oil or gel, which makes them very difficult to handle.

[0010] Therefore, the general handling of these substances is inefficient and time-consuming. Furthermore, due to the difficulty in weighing and transferring such substances and the high probability of material loss, the repeatability of any operation is poor.

[0011] Purpose of the invention In view of this, the first object of this disclosure is to provide a solid metal complex salt that does not exhibit one or more of the above-mentioned disadvantages.

[0012] Another object of this disclosure is to provide a method for generating such a solid metal complex salt.

[0013] Another object of this disclosure is to provide the use of such solid metal complex salts in the synthesis of smegglutinin, telpopritin and their pharmaceutically acceptable salts. Summary of the Invention

[0014] In a first aspect, this disclosure relates to a solid metal complex salt, which can be obtained by reacting a compound of formula I with a metal salt of formula MX: R1-(AEEA)m-A1-CO-(CH2)n-COO-R2 (I), in The amount of MX metal salt used is in the range of 0.5 to 5.0 molar equivalents relative to the compound of formula I; M is a metal cation selected from alkaline earth metal cations and transition metal cations; X is an organic or inorganic anion; R1 is selected from OH, protected or unprotected amino acids, and peptide groups consisting of 2 to 4 protected or unprotected amino acids; R2 is an H or carboxyl protecting group; A1 is a protected or unprotected amino acid, or a peptide group consisting of 2 to 4 protected or unprotected amino acids; AEEA is 2-[2-(2-aminoethoxy)ethoxy]acetyl; m is an integer between 1 and 10; and n is an integer between 8 and 28.

[0015] The reaction of compound I with MX metal salt is a complexation reaction, also known as a complex formation reaction, which is a reaction that forms a metal complex salt.

[0016] The term "metal salt," such as the MX metal salt of this disclosure, refers to any compound containing metal and non-metal atoms bonded together by ionic bonds.

[0017] The term "metal complex salt," also known as a coordination compound, refers to a complex in which a central metal ion is surrounded by a certain number of ligands (atoms, ions, or molecules that donate electrons to the metal ion to form a coordinate covalent bond). The central metal ion is usually a transition metal ion or an alkaline earth metal cation.

[0018] The term “between X and Y” is used throughout this disclosure to define the range from X to Y, including the endpoints X and Y.

[0019] The term "protecting group" refers to a temporarily attached group used to reduce the reactivity of a functional group, so that the protected functional group does not react under the synthetic conditions in one or more subsequent steps.

[0020] The term "active pharmaceutical ingredient" refers to those substances in a medicine that are responsible for the beneficial health effects experienced by consumers. An example of an API is acetaminophen, found in painkiller tablets.

[0021] Advantageously, this solid metal complex salt does not stick to the container, thus allowing for easy handling without significant loss of the compound. Furthermore, this solid metal complex salt can be readily separated, for example, in powder form, by precipitation from the solvent using a suitable antisolvent, or by removing the solvent, for example, by evaporation.

[0022] As a non-limiting example, this solid metal complex salt can therefore be readily weighed, purified, characterized, transferred to another container, stored, or used as an intermediate for further reactions without significant material loss.

[0023] On the other hand, this disclosure relates to a method for preparing a solid metal complex salt by reacting a compound of formula I with an MX metal salt: R1-(AEEA)m-A1-CO-(CH2)n-COO-R2 (I), in The amount of MX metal salt used is in the range of 0.5 to 5.0 molar equivalents relative to the compound of formula I; M is a metal cation selected from alkaline earth metal cations and transition metal cations; X is an organic or inorganic anion; R1 is selected from OH, protected or unprotected amino acids, and peptide groups consisting of 2 to 4 protected or unprotected amino acids; R2 is an H or carboxyl protecting group; A1 is a protected or unprotected amino acid, or a peptide group consisting of 2 to 4 protected or unprotected amino acids; AEEA is 2-[2-(2-aminoethoxy)ethoxy]acetyl; m is an integer between 1 and 10; and n is an integer between 8 and 28. The method includes the following steps: (i) Disperse or dissolve the compound of formula I in a first equal part of a liquid medium; (ii) Treating compound I with an MX metal salt to form a metal complex salt, and (iii) Recover metal complex salts in solid form.

[0024] The phrase "treating a compound of formula I with an MX metal salt to form a metal complex salt" refers to contacting a compound of formula I with an MX metal salt to react and form a metal complex salt.

[0025] The term "dispersion" refers to the discrete phase distribution of a compound of formula I in a liquid medium, such as consisting of particles or droplets.

[0026] Advantageously, this method converts the Formula I compound into a solid metal complex salt, which is more manageable because it is non-adhesive, i.e., does not adhere to the surface of equipment used to prepare, store or handle the compound.

[0027] This method can yield solid metal complex salts in high yield and with high purity. In fact, any impurities that are soluble in the liquid medium but cannot react with MX metal salts to form a solid will remain in the liquid phase and can therefore be easily separated from the solid metal complex salts, for example, by filtration.

[0028] Furthermore, this method requires mild conditions and involves simple and industrially feasible operations.

[0029] In this disclosure, the terms "side chain" and "lateral chain" are used interchangeably to refer to chemical groups attached to the core part of a molecule, which is called the "main chain" or backbone.

[0030] On the other hand, this disclosure relates to the use of such solid metal complex salts in the synthesis of smegglutinin, telpopritin and their pharmaceutically acceptable salts.

[0031] Advantageously, the use of this solid metal complex salt makes the synthesis of smegglutinin, telpopritin, and their pharmaceutically acceptable salts highly efficient, simple, and industrially feasible.

[0032] In addition, it reduces the loss of expensive intermediates (such as the intermediates in Formula I).

[0033] Furthermore, the apparatus for synthesizing smegglutide, telpolide, and their pharmaceutically acceptable salts is faster and easier to clean.

[0034] Compared with synthetic methods involving the use of viscous compounds of formula I without complexation, the method according to the invention can obtain smegglutinin, telpoxetine and their pharmaceutically acceptable salts in higher yields and purities.

[0035] On the other hand, this disclosure relates to a method for preparing an active pharmaceutical ingredient, said active pharmaceutical ingredient including smegglutinin, telpoxetine and pharmaceutically acceptable salts thereof, characterized in that the method uses said solid metal complex salt. Attached Figure Description

[0036] Some features and advantages of the present invention will become apparent from the accompanying drawings, in which: Figure 1 This is a schematic diagram of possible compounds of Formula 1, where R1, R2, A1 and AEEA have been described; Figure 2 The HPLC-MS chromatogram of the solid metal complex salt prepared in Example 2 is shown. Figure 3 The HPLC-MS chromatogram of the solid metal complex salt prepared in Example 4 is shown. Figure 4 The HPLC-MS chromatograms of smegglutinin fragment 20-31 obtained by linking the free side chain fragment with fragment 21-31 are shown (upper part of the figure), and the HPLC-MS chromatograms of the same fragment obtained by linking the complexed side chain fragment obtained according to the present invention with fragment 21-31 are shown (lower part). Figure 5 The HPLC-MS chromatograms of the telpoeptide 20-29 fragment obtained by linking the free side chain fragment with the 21-29 fragment are shown (upper part of the figure), and the HPLC-MS chromatograms of the same fragment obtained by linking the complexed side chain fragment obtained according to the present invention with the 21-29 fragment are shown (lower part). Figure 6 The following are shown: compound (F1) of Formula 1 (6a), the metal complex salt (6b) obtained by reacting the compound of Formula 1 with 1 molar equivalent of MgCl2, and the metal complex salt (6c) obtained by reacting the compound of Formula 1 with 2 molar equivalents of MgCl2. 1 H-NMR spectra. The figure highlights the proton signal positions of the NH group belonging to Glu (marked with a triangle Δ), the two AEEA groups (marked with squares or circles respectively), the α-NH group of Lys (marked with a rhombus), and the ε-NH group (marked with a triangle at the head).

[0037] Figure 7 The spatial arrangement of the compounds of Formula 1 is shown, calculated based on the presence and amount of MgCl2 used. Detailed Implementation

[0038] In some embodiments, the metal cation M is selected from the group consisting of Ca, Zn, Mg, Cu, Co, Ni, Ti, Zr, Fe, and Ag cations. Preferably, the metal cation is selected from Ca. 2+ Zn 2+ Mg 2+ Cu 2+ Co 2+ Ni 2+ Ti 2+ Zr 2+ Fe 3+ and Ag + The group consisting of, more preferably, is selected from Cu 2+ Fe 3+ Co 2+ Mg 2+ The group consisting of, or even more preferably, the metal cation M is Mg 2+ or Cu 2 + Most preferably, the metal cation M is Mg. 2+ .

[0039] In some embodiments, X is a halide anion, preferably X is Cl. -1 or Br -1 More preferably, X is Cl -1 .

[0040] In some embodiments, the MX metal salt is MgCl2 or CuCl2, and in some preferred embodiments, the MX metal salt is MgCl2.

[0041] In some embodiments, the amount of the MX metal salt is in the range of 0.8 to 4.0 molar equivalents relative to the compound of formula I, preferably in the range of 1.0 to 2.5 molar equivalents.

[0042] In some preferred embodiments, the MX metal salt is MgCl2, and its amount is in the range of 0.8 to 4.0 molar equivalents relative to the compound of formula I, preferably in the range of 1.0 to 2.5 molar equivalents.

[0043] In the reaction, increasing the amount of MX metal salt relative to the compound of formula I promotes the formation of solid metal complex salts.

[0044] However, when the MX metal salt is used in a low molar equivalent relative to the compound of Formula I (where low means in the range of 1.0 to 2.5), the stability of the solid metal complex salt is improved compared to the stability of the solid metal complex salt obtained in a high molar equivalent (where high means in the range of 3.0 to 5.0). In particular, when the solid metal complex salt is obtained in a low molar equivalent relative to the compound of Formula I, the undesirable removal of one or more carboxyl protecting groups due to the HCl generated during the formation of the solid metal complex salt is reduced.

[0045] Solid metal complex salts obtained with high molar equivalents of MX metal salts compared to compounds of Formula I exhibit slight viscosity. Furthermore, the hygroscopicity of solid metal complex salts obtained with low molar equivalents of MX metal salts compared to compounds of Formula I is reduced. Additionally, solid metal complex salts obtained with low molar equivalents of MX metal salts compared to compounds of Formula I are easier to filter. This is presumably due to their larger particle size.

[0046] In some embodiments, R1 is protected or unprotected Lys and / or A1 is protected or unprotected Glu; in some preferred embodiments, R1 is protected or unprotected Lys and A1 is protected or unprotected Glu; in some more preferred embodiments, R1 is protected Lys and A1 is protected Glu.

[0047] In some other embodiments, R1 is OH and / or Al is protected or unprotected Glu; in some preferred embodiments, R1 is OH and Al is protected or unprotected Glu; in some more preferred embodiments, R1 is OH and Al is protected Glu.

[0048] In some embodiments, the compound of formula I has one or more carboxyl groups protected by a protecting group, the protecting group being independently selected from tert-butyl (tBu), β-3-methylpentan-3-yl (Mpe), 2-phenylisopropyl (2-Ph'Pr) and 4-(3,6,9-trioxaundecyl)oxybenzyl (TEGBz or TEGBn).

[0049] In some embodiments, A1 is protected Glu, and the α-carboxyl group of Glu is preferably protected by a tert-butyl protecting group.

[0050] In some embodiments, all protecting groups of all carboxyl groups in the compound of formula I are tert-butyl protecting groups.

[0051] In some embodiments, the Formula I compound has one or more amino groups protected by a protecting group, the protecting group being independently selected from Fmoc, Boc, Smoc, and Bz.

[0052] In some embodiments, when R1 is an amino-protected amino acid, the amino protecting group of R1 is selected from the group consisting of Fmoc, Boc, Smoc and Bz.

[0053] In some embodiments, R1 is a protected Lys, the α-amino group of which is preferably protected by an Fmoc protecting group.

[0054] In some embodiments, R2 is a carboxyl protecting group, preferably tert-butyl.

[0055] In some preferred embodiments, A1 is protected Glu, the α-carboxyl group of which is protected by a tert-butyl protecting group; R1 is protected Lys, the α-amino group of which is protected by a Fmoc protecting group.

[0056] According to some implementation schemes, m is an integer between 1 and 5, preferably between 1 and 4, and more preferably m is 2.

[0057] According to some implementation schemes, n is an integer between 10 and 25; preferably, n is between 15 and 21.

[0058] According to some preferred embodiments, m is contained between 1 and 4, and n is contained between 15 and 21, more preferably m is 2, and n is contained between 15 and 21.

[0059] AEEA is preferably linked to R1 via the C atom of the ketone group, and / or AEEA is preferably linked to Al via the N atom of the amino group, and / or when there is more than one AEEA, they are preferably linked to each other via amide bonds.

[0060] When m is 2 and both A1 and R1 are selected from protected or unprotected amino acids or peptide groups composed of 2 to 4 protected or unprotected amino acids, one side of an AEEA is connected to A1 via an amide link, so that the N atom of the amino group of the AEEA is covalently bonded to the C atom of the ketone group of A1, and the other side is connected to another AEEA via an amide link, so that the C atom of the ketone group of the first AEEA is covalently bonded to the N atom of the amino group of the second AEEA; the second AEEA is connected to R1 via an amide link, so that the C atom of the ketone group of the second AEEA is covalently bonded to the N atom of the amino group of R1.

[0061] In some embodiments, when m is 2, A1 is protected or unprotected Glu, and R1 is protected or unprotected Lys, one side of an AEEA is connected to Glu via an amide link, such that the N atom of the amino group of the AEEA is covalently bonded to the C atom (δC) of the ketone group of Glu, and the other side is connected to another AEEA via an amide link, such that the C atom of the ketone group of the first AEEA is covalently bonded to the N atom of the amino group of the second AEEA, and the second AEEA is connected to Lys via an amide link, such that the C atom of the ketone group of the second AEEA is covalently bonded to the εN atom of Lys.

[0062] The present invention includes all different embodiments that can be obtained by combining features that define m as an integer between 1 and 5 (preferably between 1 and 4, more preferably m is 2) and features that define n as an integer between 10 and 25 (between 15 and 21). These features can also be freely combined with features of each embodiment disclosed above with respect to different proportions of metal cations, halide anions, R1, R2, Al, and MX metal salts with compounds of formula I.

[0063] The terms “proportion” and “equivalence ratio” refer to the number of molar equivalents of the MX metal salt used relative to the number of molar equivalents of the compound of formula I.

[0064] In some embodiments, the compound of formula I is a compound of formula 1: (1) Or compounds of formula 2: (2) Or compounds of formula 3: (3) Or compounds of formula 4: (4).

[0065] Figure 1 Compounds of Formula 1 are reported, with R1, R2, A1, and AEEA groups illustrated as possible examples.

[0066] In some preferred embodiments, the MX metal salt is MgCl2, and the amount of the MX metal salt is in the range of 0.5 to 5.0 molar equivalents relative to the compound of formula I, preferably in the range of 1.0 to 2.5; R1 is protected Lys or OH; R2 is a carboxyl protecting group; A1 is protected Glu; m is 2 and n is 16 or 18. In some more preferred embodiments, the MX metal salt is MgCl2, and the amount of the MX metal salt is in the range of 0.5 to 5.0 molar equivalents relative to the compound of formula I, preferably in the range of 1.0 to 2.5; R1 is Fmoc protected Lys or OH; R2 is tert-butyl; A1 is tert-butyl protected Glu; m is 2 and n is 16 or 18.

[0067] In some preferred embodiments, the MX metal salt is MgCl2, the amount of the MX metal salt is in the range of 1.0 to 2.5 molar equivalents relative to the compound of formula I, R1 is Fmoc-protected Lys or OH, R2 is tert-butyl, A1 is tert-butyl-protected Glu, m is 2 and n is 16 or 18.

[0068] In a particularly preferred embodiment, the MX metal salt is MgCl2, the amount of the MX metal salt is in the range of 1.0 to 2.5 molar equivalents relative to the compound of formula I, R1 is Fmoc-protected Lys or OH, R2 is tert-butyl, A1 is tert-butyl-protected Glu, m is 2 and n is 16. The embodiment in which R1 is Fmoc-protected Lys is even more preferred.

[0069] In another particularly preferred embodiment, the MX metal salt is MgCl2, the amount of the MX metal salt is in the range of 1.0 to 2.5 molar equivalents relative to the compound of formula I, R1 is Fmoc-protected Lys or OH, R2 is tert-butyl, A1 is tert-butyl-protected Glu, m is 2 and n is 18. The embodiment in which R1 is Fmoc-protected Lys is even more preferred.

[0070] The adhesive properties of compounds of Formula I, especially those of Formulas 1 to 4, can be reasonably attributed to the coexistence of the hydrophilic AEEA moiety and the alkylated chain moiety.

[0071] This article also provides a method for preparing the above-mentioned solid metal complex salt, the method comprising the step of treating the compound of formula I with an MX metal salt to form a metal complex salt, wherein the compound of formula I is dispersed or dissolved in a first fraction of a liquid medium beforehand, and the step of recovering the metal complex salt in solid form.

[0072] According to some implementation schemes, step iii) of recovering the solid form of the metal complex salt includes antisolvent precipitation or removal of the liquid medium, preferably including antisolvent precipitation.

[0073] Advantageously, antisolvent precipitation is a simple and flexible technique; in fact, precipitation can be easily optimized by selecting the solvent and antisolvent, their amounts, their ratios, and the operating temperature.

[0074] Furthermore, antisolvent precipitation can be implemented industrially without expensive or complex equipment.

[0075] The liquid medium may be selected from the group consisting of acetone, water, methanol, isopropanol, ethanol, dichloromethane, chloroform, ethyl acetate, methyl acetate, isopropyl acetate, acetonitrile, dimethyl carbonate, toluene, tetrahydrofuran, and mixtures thereof.

[0076] In step (i), the compound of formula I can be dispersed or dissolved in a liquid medium; preferably, it is dissolved in a liquid medium.

[0077] Advantageously, when a liquid medium capable of dissolving Formula I compound is used in step (i), there is no loss due to Formula I compound eventually adhering to the surface of the container or vessel.

[0078] Furthermore, the formation kinetics of the metal complex salt are improved when the compound of formula I is dissolved in a liquid medium compared to when the compound of formula I is dispersed in a liquid medium; in other words, the rate of obtaining the metal complex salt is increased when the compound of formula I is dissolved in a liquid medium compared to when the compound of formula I is dispersed in a liquid medium.

[0079] According to some embodiments, the liquid medium is selected from the group consisting of acetone, ethyl acetate, dimethyl carbonate, and mixtures of dichloromethane and tetrahydrofuran; preferably, the mixture of dichloromethane and tetrahydrofuran is in the range of 5:1 to 15:1 (vol:vol), and even more preferably in the range of 8:1 to 10:1 (vol:vol).

[0080] According to some preferred embodiments, the liquid medium is acetone, ethyl acetate or dimethyl carbonate, and according to some more preferred embodiments, the liquid medium is acetone.

[0081] Dimethyl carbonate is a well-established solvent and green reagent, and has been attracting much attention due to its favorable biodegradability in the atmosphere and non-toxicity.

[0082] In some embodiments, the solid metal complex salt is recovered by precipitation with an antisolvent selected from the group consisting of heptane, toluene, acetonitrile, ethyl acetate, cyclopentylmethyl ether, petroleum ether, diisopropyl ether, methyl tert-butyl ether, and mixtures thereof. Preferably, the antisolvent is selected from the group consisting of ethyl acetate, diisopropyl ether, and mixtures thereof.

[0083] In a preferred embodiment of the above-described method for preparing solid metal complex salts, the solid metal complex salts are recovered by antisolvent precipitation. The liquid medium is acetone, ethyl acetate, or dimethyl carbonate, and the antisolvent is acetone, ethyl acetate, or isopropyl ether, or a mixture of acetone and isopropyl ether, or a mixture of ethyl acetate and isopropyl ether.

[0084] In some implementations, the liquid medium and the antisolvent are the same chemical or a mixture of the same chemicals.

[0085] In other words, the liquid medium in step (i) for dispersing or dissolving compound I is an antisolvent for the metal complex salt. This improves the overall efficiency of the method in terms of time, reduces the use of different chemical products, and makes the recovery of this liquid medium easier.

[0086] In some embodiments, step (ii) is performed by adding the metal salt MX to the dispersion or solution produced in step (i), wherein the MX metal salt is not previously dispersed or dissolved in any liquid medium; preferably, the compound of formula I is previously dissolved in a first aliquot of a liquid medium, and the MX metal salt is added directly to that solution.

[0087] In an alternative embodiment, step (ii) is performed by adding the MX metal salt to the dispersion or solution generated in step (i), or by adding the dispersion or solution generated in step (i) to the MX metal salt, wherein the MX metal salt is previously dispersed or dissolved in a second portion of the liquid medium.

[0088] In some embodiments, step (ii) of treating the compound of formula I with an MX metal salt to form a metal complex salt is carried out in the presence of a buffer (preferably an alkaline buffer, more preferably selected from the group consisting of ammonium acetate, sodium ascorbate, sodium tetraborate, potassium tetraborate and K-Oxyma) or a weak base (preferably any tertiary amine, more preferably triethylamine).

[0089] The term "alkaline buffer" refers to substances that can maintain an alkaline pH in a solution.

[0090] Advantageously, in the presence of an alkaline buffer, the compound of formula I is treated with an MX metal salt to form a metal complex salt, which makes the metal complex salt more stable.

[0091] In some preferred embodiments, an alkaline buffer or weak base is added to the dispersion or solution of the compound of formula I in step (i) in the first aliquot of the liquid medium.

[0092] According to some implementation schemes, step iii) of recovering the solid form of the metal complex salt includes antisolvent precipitation followed by filtration.

[0093] This results in a high recovery rate for solid metal complex salts.

[0094] According to some implementation schemes, step iii) of recovering the solid form of the metal complex salt includes antisolvent precipitation, wherein both the liquid medium and the antisolvent are ethyl acetate.

[0095] In these embodiments, ethyl acetate serves as both a solvent for the compound of formula I and an antisolvent for the metal complex salt.

[0096] According to some alternative embodiments, step iii) of recovering the metal complex salt in solid form includes antisolvent precipitation, wherein the liquid medium is ethyl acetate and the antisolvent is a mixture of ethyl acetate and diisopropyl ether.

[0097] Furthermore, in such embodiments, ethyl acetate serves as both a solvent for the compound of formula I and an antisolvent for the metal complex salt; however, in these cases, diisopropyl ether is also added as an additional antisolvent for the metal complex salt.

[0098] Advantageously, in these embodiments, the addition of diisopropyl ether accelerates the precipitation of solid metal complex salts and / or yields free-flowing, powdery solid precipitates of the metal complex salts, especially when the equivalence ratio of the MX metal salt to the compound of formula I is low. Furthermore, the addition of diisopropyl ether improves the yield.

[0099] If the powder particles are not stuck together, the powder is defined as a free-flowing powder.

[0100] According to some other alternative embodiments, step iii) of recovering the metal complex salt in solid form includes antisolvent precipitation, wherein the liquid medium is dimethyl carbonate and the antisolvent is ethyl acetate and / or diisopropyl ether.

[0101] In addition, in this case, the use of diisopropyl ether in addition to ethyl acetate will accelerate the precipitation of solid metal complex salts and promote the formation of solid metal complex salts at low molar equivalents, i.e. when the amount of MX metal salt is in the range of 1.0 to 2.5 molar equivalents relative to the compound of formula I.

[0102] According to some preferred alternative embodiments, step iii) of recovering the solid form of the metal complex salt from the liquid medium includes antisolvent precipitation, wherein the liquid medium is acetone and the antisolvent is acetone and / or diisopropyl ether.

[0103] The dispersion or dissolution of the compound of formula I in the first fraction of the liquid medium in step (i) is preferably carried out at room temperature, more preferably in a temperature range of about 20°C to 30°C, and even more preferably in a temperature range of about 20°C to 25°C.

[0104] The term "room temperature" refers to a temperature range of approximately 15°C to 35°C.

[0105] Preferably, step (ii) is also carried out at room temperature, more preferably in a temperature range of about 20°C to 30°C, and even more preferably in a temperature range of about 20°C to 25°C.

[0106] In some embodiments, after step (ii), the temperature is reduced to a range between 0°C and 15°C, preferably between 0°C and 10°C or between 10°C and 15°C.

[0107] Advantageously, this can accelerate the precipitation of solid metal complex salts.

[0108] In some implementations, when the liquid medium is not an antisolvent for the metal complex salt, its amount can be reduced before the antisolvent is added, for example by partial evaporation, so that a small amount of antisolvent is used to precipitate the solid metal complex salt.

[0109] In some alternative embodiments, precipitation of solid metal complex salts is achieved by removing the liquid medium, for example by evaporation, freeze drying, spray drying, permeate evaporation membrane technology, or equivalent techniques.

[0110] This allows for the recovery of at least a portion of the liquid medium.

[0111] In some implementations, the solid metal complex salt is washed after filtration.

[0112] Advantageously, this reduces impurities in solid metal complex salts.

[0113] In some preferred embodiments, the solid metal complex salt is washed with diisopropyl ether after filtration.

[0114] When using ethyl acetate as the liquid medium, the resulting solid metal complex salt is preferably washed once or multiple times with a mixture of diisopropyl ether and / or acetonitrile and heptane.

[0115] A mixture of acetonitrile and heptane can be used to remove ethyl acetate that may remain in solid metal complex salts.

[0116] In some embodiments, the solid metal complex salt is dried after washing, preferably under nitrogen.

[0117] In some preferred embodiments, after washing, the solid metal complex salt is dried at a temperature range of 0°C to 15°C, preferably 0°C to 10°C or 10°C to 15°C, preferably under nitrogen.

[0118] The invention also provides the use of solid metal complex salts of the present invention in the synthesis of active pharmaceutical ingredients, more preferably the use of solid metal complex salts of compounds of formulas 1 to 4 in the synthesis of active pharmaceutical ingredients. Furthermore, the invention also provides the use of solid metal complex salts of the present invention in the synthesis of smegglutide, telpolide, and pharmaceutically acceptable salts thereof. Most preferably, the use of solid metal complex salts of compounds of formulas 1 to 4 in the synthesis of smegglutide, telpolide, and pharmaceutically acceptable salts thereof is provided.

[0119] A method for synthesizing active pharmaceutical ingredients using the solid metal complex salt of the present invention is also provided. More preferably, the present invention includes a method for preparing smegglutinin, telpoxetine, and pharmaceutically acceptable salts thereof, characterized by using the solid metal complex salt disclosed in the present invention.

[0120] In some embodiments, the synthesis of smegglutide, telpolide, and pharmaceutically acceptable salts thereof includes the preparation of solid metal complex salts of compounds of formula I, preferably solid metal complex salts of compounds of formulas 1-4, and more preferably solid metal complex salts of compounds of formulas 1 and 2.

[0121] In fact, compounds of Formula I and solid metal complex salts of Formulas 1-4 can be used as intermediates in the synthesis of smegglutinin, telposide and their pharmaceutically acceptable salts.

[0122] In particular, in the synthesis of smegglutinin, telpokinin and their pharmaceutically acceptable salts, the solid metal complex salts of Formula I compounds can be used as is for coupling reactions: the amino acid or peptide fragments of the solid metal complex salts of Formula I can be directly coupled with other amino acids or peptide fragments without prior decomplexing steps.

[0123] As a non-limiting example, the solid metal complex salt of compound 1 can be used as is for coupling reaction with fragments 21-31 of smegglutinin to generate fragments 20-31 of smegglutinin; the solid metal complex salt of compound 2 can also be used directly for coupling reaction with fragments 21-29 of telposide to generate fragments 20-29.

[0124] Similarly, the solid metal complex salts of Formulas 3 and 4 can be activated and coupled with other amino acids or peptide fragments to generate fragments of smegglutinin and telposide, respectively.

[0125] Experimental Section Example 1 - Synthesis and isolation of iron (III) solid complex (solvent: DCM / THF) 1 g of compound 1 was dissolved in 3.6 mL of a first mixed solvent of DCM / THF with a volume ratio of 9:1 at room temperature to obtain a DCM / THF solution of compound 1 containing 280 g of compound 1 per liter of a second mixed solvent.

[0126] 0.54 g of anhydrous FeCl3 (3.0 molar equivalents relative to the compound of Formula 1) was dissolved in 21.7 mL of a second mixed solvent of DCM / THF with a volume ratio of 9:1 at room temperature and kept under stirring for 30 minutes to obtain a DCM / THF solution of FeCl3.

[0127] Then, the DCM / THF solution of FeCl3 and the DCM / THF solution of compound 1 were mixed at room temperature and kept stirring for 30 minutes.

[0128] Finally, 10 volumes of petroleum ether were added as an antisolvent to a DCM / THF solution of FeCl3 and compound 1 at 25±5°C to precipitate the iron(III) complex salt. The volume ratio of petroleum ether used as an antisolvent to the total amount of DCM / THF used to dissolve compound 1 and FeCl3 was 6:1.

[0129] The iron(III) complex salt was then separated by centrifugation, the liquid portion was removed, and the salt was dried to obtain a brown solid powder. The powder was easy to handle because its adhesion behavior was negligible.

[0130] Example 2 - Synthesis and isolation of copper (II) solid complex (solvent: DCM / THF) 1 g of compound 1 was dissolved in 3.6 mL of a first mixed solvent of DCM / THF with a volume ratio of 9:1 at room temperature to obtain a DCM / THF solution of compound 1 containing 280 g of compound 1 per liter of a second mixed solvent.

[0131] 0.45 g of anhydrous CuCl2 (2.0 molar equivalents relative to the compound of Formula 1) was dissolved in 21.7 mL of a second mixed solvent of DCM / THF with a volume ratio of 9:1 at room temperature and kept under stirring for 30 minutes to obtain a DCM / THF solution of CuCl2.

[0132] Then, the DCM / THF solution of CuCl2 and the DCM / THF solution of compound 1 were mixed at room temperature and kept stirring for 30 minutes.

[0133] Finally, petroleum ether was added as an antisolvent to a DCM / THF solution of CuCl2 and compound of formula 1 at 25±5℃ to precipitate the copper(II) complex salt. The volume ratio of petroleum ether used as an antisolvent to the total amount of DCM / THF used to dissolve compound of formula 1 and CuCl2 was 6:1.

[0134] The copper(II) complex salt obtained therefrom is separated by filtration; a green solid hygroscopic powder is obtained; the powder is easy to transfer and weigh and does not stick to the container.

[0135] Copper(II) complex salts were analyzed using UHPLC equipped with a four-stage pump, UV detector, and ESI-Q-TOF mass spectrometer. Mobile phase A was water + 0.1% TFA, and mobile phase B was acetonitrile. Analysis demonstrated that the tert-butoxy-protected peptide underwent slight deprotection during the synthesis and / or precipitation of magnesium(II) complex salts. In fact, from... Figure 2 The HPLC-MS chromatograms of the reported solid metal complex salts show that approximately 1.5% of the tert-butyl protecting groups were removed from the peptides during the synthesis and / or precipitation of the magnesium(II) complex salts. In the chromatogram, the peaks at approximately 49 min and 55 min can be attributed to the Formula 1 compound lacking either of the two tert-butyl protecting groups and the Formula 1 compound lacking one of the two tert-butyl protecting groups, respectively, while the peak at 64 min can be attributed to the Formula 1 compound.

[0136] Example 3 - Synthesis and isolation of cobalt (II) solid complex (solvent: DCM / THF) 0.33 g of anhydrous CoCl2 was dissolved in 21.7 mL of a DCM / THF mixed solvent with a volume ratio of 9:1 at room temperature and kept stirring for 30 minutes to obtain a DCM / THF solution of CoCl2.

[0137] 1 g of compound 1 was dissolved in 3.6 mL of a DCM / THF mixed solvent with a volume ratio of 9:1 at room temperature to obtain a DCM / THF solution of compound 1 containing 280 g of compound 1 per liter of the mixed solvent.

[0138] Then, the DCM / THF solution of CoCl2 and the DCM / THF solution of compound 1 were mixed at room temperature.

[0139] Finally, MTBE was added as an antisolvent to a DCM / THF solution of CoCl2 and compound of formula 1 at 10±5 °C to precipitate the cobalt(II) complex salt. The volume ratio of MTBE used as an antisolvent to the total amount of DCM / THF used to dissolve compound of formula 1 and CoCl2 was 6:1.

[0140] The resulting cobalt(II) complex salt is separated by filtration, yielding a blue solid hygroscopic powder that is easy to transfer and weigh and does not adhere to containers.

[0141] Furthermore, precipitates of cobalt(II) complexes were successfully obtained by replacing MTBE with DIPE and petroleum ether as non-solvents. In these cases, the volume ratio of DIPE or petroleum ether used as the antisolvent to the total amount of DCM / THF used to dissolve compound 1 and CoCl2 was 0.3:1 (weight ratio).

[0142] Furthermore, in these cases, the resulting cobalt(II) complex salt is a solid, non-sticky powder.

[0143] Example 4 - Synthesis and isolation of magnesium (II) solid complex (solvent: DCM / THF) 0.32 g of anhydrous MgCl2 was dissolved in 21.7 mL of a DCM / THF mixed solvent with a volume ratio of 9:1 at room temperature and kept stirring for 30 minutes to obtain a DCM / THF solution of MgCl2.

[0144] 1 g of compound 1 was dissolved in 3.6 mL of a DCM / THF mixed solvent with a volume ratio of 9:1 at room temperature to obtain a DCM / THF solution of compound 1 containing 280 g of compound 1 per liter of the mixed solvent.

[0145] Then, the DCM / THF solution of MgCl2 and the DCM / THF solution of compound 1 were mixed at room temperature so that the weight ratio of MgCl2 to compound 1 was 0.3:1 (by weight).

[0146] Finally, MTBE was added as an antisolvent to a DCM / THF solution of MgCl2 and compound of formula 1 at 10±5 °C to precipitate the magnesium(II) complex salt. The volume ratio of MTBE used as an antisolvent to the total amount of DCM / THF used to dissolve compound of formula 1 and MgCl2 was 0.3:1 (by weight).

[0147] The resulting magnesium(II) complex salt is obtained by filtration; it is a green, hygroscopic powder that can be handled, transferred, and weighed very easily without material loss on the container walls.

[0148] Magnesium(II) complex salts were analyzed using UHPLC equipped with a four-stage pump, UV detector, and ESI-Q-TOF mass spectrometer. Mobile phase A consisted of water + 0.1% TFA, and mobile phase B consisted of acetonitrile. Analysis demonstrated that the tert-butyl-protected peptides did not undergo deprotection during the synthesis and precipitation of the magnesium(II) complex salts. In fact, from... Figure 3 According to the LC-MS spectrum of the solid metal complex salt in the report, only the peak at 64 minutes, which belongs to the compound of formula 1, is visible.

[0149] Example 5 - Synthesis of semaglutide fragment 20-31 - attachment of side chains According to the SPPS method, 1.3g of the magnesium(II) complex from Example 4 was coupled with fragments 21-31 of smegglutinin to obtain fragments 20-31 of smegglutinin.

[0150] First, the powdered solid metal complex salt is dissolved in an organic solvent (DMF, DCM), and then 130 µL of diisopropylcarbodiimide and 118 mg of OxymaPure® are added for activation to generate a coupling mixture.

[0151] The coupling mixture was then added to swollen L-Glu(PG)-L-Phe-L-Ile-L-Ala-L-Trp(PG)-L-Leu-L-Val-L-Arg(PG)-Gly-L-Arg(Pbf)-Gly-MBH resin (fragments 21-31, protected and attached to MBH resin), equivalent to 2 g of starting resin, with a loading of 0.4 mmol / g; the reaction was carried out at 40 °C for 8 hours.

[0152] In this specification, PG represents a protecting group. In Example 5, the PG of Glu and Trp is tBU, and the PG of Arg is Pbf; therefore, the resin L-Glu(PG)-L-Phe-L-Ile-L-Ala-L-Trp(PG)-L-Leu-L-Val-L-Arg(PG)-Gly-L-Arg(Pbf)-Gly-MBH is L-Glu(tBU)-L-Phe-L-Ile-L-Ala-L-Trp(tBU)-L-Leu-L-Val-L-Arg(Pbf)-Gly-L-Arg(Pbf)-Gly-MBH is a resin L-Glu(tBU)-L-Phe-L-Ile-L-Ala-L-Trp(tBU)-L-Leu-L-Val-L-Arg(Pbf)-Gly-L-Arg(Pbf)-Gly-MBH.

[0153] The mixture was then filtered, and the resin was washed with fresh organic solvent. Coupling efficiency was assessed by colorimetric tests (Kaiser test, TNBS test, and Chloranil test), and no unreacted sites were found.

[0154] The coupling efficiency can also be detected by cleavage in a mixture of TFA / DTT / TIS / water (87.5 / 5 / 5 / 2.5 v / w / v / v) followed by HPLC analysis. Figure 4 As shown, the figure reports the HPLC chromatograms of fragment 20-31 of smegglutinin. Specifically, the upper figure refers to the unprotected fragment 20-31 obtained by coupling the free (i.e., uncomplexed) side chain to fragment 21-31, while the lower figure refers to the unprotected fragment 20-31 obtained by coupling the side chain, which has been previously complexed by reaction with MgCl2 metal salt, to fragment 21-31.

[0155] It can be clearly seen that, with the use of complexed side chains, coupling with fragments 21-31 was also successfully achieved.

[0156] Example 6 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) 15g of compound 1 was dissolved in 75mL of ethyl acetate at room temperature to obtain an ethyl acetate solution of compound 1.

[0157] 1.8 g of anhydrous MgCl2 was added to 75 mL of ethyl acetate at room temperature and stirred at 20-30 °C for 30 minutes; then an ethyl acetate solution of compound 1 was added and stirred at 20-30 °C for 10 minutes, and then cooled to 0-10 °C to precipitate magnesium(II) complex salt.

[0158] The solid magnesium(II) complex salt was separated by filtration, washed twice with 30 mL of ethyl acetate, and then dried under nitrogen and then under vacuum.

[0159] Example 7 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetonitrile) 1.0 g of compound 1 was dissolved in 32 mL of acetonitrile at room temperature and stirred until a clear solution was obtained. Then, 320 mg of MgCl2 (4.0 molar equivalents relative to compound 1) was added to the solution, and the solid metal complex salt immediately began to precipitate.

[0160] Stir the mixture at 20-30°C for 120 minutes, then cool it to 0-10°C and stir it again for 120 minutes.

[0161] The solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, and dried under nitrogen at 0-10 °C for 60 minutes.

[0162] Yield: 1.25g solid metal complex salt.

[0163] Example 8 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetonitrile) 1.0 g of compound 1 was dissolved in 32.5 mL of acetonitrile and stirred until a clear solution was obtained; then 240 mg of MgCl2 (3.0 molar equivalent relative to compound 1) was added to the solution, and the metal complex salt began to form.

[0164] Stir the mixture at 25°C for 30 minutes, then cool it to 0-10°C and stir it again for 120 minutes.

[0165] Add 125 mL of diisopropyl ether to the mixture and stir at 0-10 °C for 120 minutes.

[0166] Finally, the solid metal complex salt was separated by filtration, washed with 5 mL of diisopropyl ether, and dried under nitrogen at 0-10 °C for 120 minutes.

[0167] Yield: 1.20g solid metal complex salt.

[0168] Example 9 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 20g of compound 1 in 100mL of ethyl acetate and stir until a clear solution is obtained.

[0169] 2.40 g of MgCl2 (1.5 molar equivalents relative to compound 1) was added to 100 mL of ethyl acetate and stirred at 20-30 °C for 30 minutes. Then, an ethyl acetate solution of compound 1 was added, and the solid metal complex salt immediately began to precipitate. Stirring was continued at 20-30 °C for 10 minutes. The mixture was then cooled to 0-10 °C and stirred for another 180 minutes.

[0170] Finally, the solid metal complex salt was separated by filtration, washed with three equal portions of ethyl acetate (3 × 20 mL), dried under nitrogen for 30 minutes, and then dried under vacuum for 5 hours, with the temperature maintained between 0°C and 10°C throughout.

[0171] Yield: 21.00g solid metal complex salt.

[0172] Example 10 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0173] When 320 mg of MgCl2 (4.0 molar equivalents relative to compound 1) is added to the solution, the solid metal complex salt immediately begins to precipitate.

[0174] Stir the mixture at 20-30°C for 120 minutes, then cool it to 0-10°C and keep it stirred for another 120 minutes.

[0175] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of ethyl acetate, dried under nitrogen for 60 minutes, and dried under vacuum overnight, with the temperature maintained between 0°C and 10°C throughout.

[0176] Yield: 1.30g solid metal complex salt.

[0177] Example 11 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate with stirring until a clear solution is obtained.

[0178] When 80 mg of MgCl2 (1.0 molar equivalent relative to compound 1) is added to the solution, a metal complex salt begins to form.

[0179] Stir the mixture at 20-30°C for 120 minutes.

[0180] Then add 15 mL of diisopropyl ether, stir overnight at 20-30°C, cool to 0-10°C and keep stirring for another 120 minutes.

[0181] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of ethyl acetate, and dried in a vacuum overnight, with the temperature maintained between 0°C and 5°C.

[0182] Yield: 1.00g solid metal complex salt.

[0183] Example 12 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0184] 88 mg of MgCl2 (1.1 molar equivalents relative to compound 1) was added to 5 mL of ethyl acetate. Then, the ethyl acetate solution of compound 1 was added in two portions according to the following steps: first, 25% of the volume of the solution was added at 25°C and maintained at 25°C with stirring for 30 minutes; then, the remaining 75% of the volume of the solution was added at 25°C and maintained at 25°C with stirring for another 30 minutes. During the addition of the ethyl acetate solution of compound 1, a metal complex salt began to form.

[0185] Then add 25 mL of diisopropyl ether, cool the mixture to 0-10 °C and keep it stirred for 120 minutes.

[0186] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, and dried under nitrogen for 4 hours, with the temperature maintained between 0°C and 10°C.

[0187] Yield: 1.00g solid metal complex salt.

[0188] Example 13 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0189] When 100 mg of MgCl2 (1.2 molar equivalents relative to compound 1) is added to the solution, a metal complex salt begins to form.

[0190] Stir the mixture at 20-30°C for 120 minutes.

[0191] Then add 15 mL of diisopropyl ether, cool the mixture to 0-10 °C and keep it stirred for 120 minutes.

[0192] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, dried under nitrogen for 60 minutes, and then dried under vacuum overnight, with the temperature maintained between 0°C and 5°C.

[0193] Yield: 1.05g solid metal complex salt.

[0194] Example 14 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0195] When 120 mg of MgCl2 (1.5 molar equivalents relative to compound 1) is added to the solution, the solid metal complex salt immediately begins to precipitate.

[0196] The mixture was kept at 20-30°C with stirring for 120 minutes; then 15 mL of diisopropyl ether was added, the mixture was cooled to 0-10°C and kept with stirring for another 120 minutes.

[0197] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, dried under nitrogen for 1 hour, and then dried under vacuum overnight, with the temperature maintained between 0°C and 5°C.

[0198] Yield: 1.10g solid metal complex salt.

[0199] Example 15 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0200] When 160 mg of MgCl2 (2.0 molar equivalents relative to compound 1) is added to the solution, the solid metal complex salt immediately begins to precipitate.

[0201] The mixture is kept at 20-30°C with stirring for 120 minutes, then cooled to 0-10°C and kept with stirring for another 120 minutes.

[0202] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, and dried under nitrogen at 0°C to 5°C for 60 minutes.

[0203] Yield: 1.10g solid metal complex salt.

[0204] Example 16 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0205] When 240 mg of MgCl2 (3.0 molar equivalents relative to compound 1) is added to the solution, the solid metal complex salt immediately begins to precipitate.

[0206] The mixture is kept at 20-30°C with stirring for 120 minutes, then cooled to 0-10°C and kept with stirring for another 120 minutes.

[0207] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, and dried under nitrogen at 0°C to 10°C for 1 hour.

[0208] Yield: 1.15g solid metal complex salt.

[0209] Example 17 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 1.0 g of compound 1 in 5 mL of ethyl acetate and stir until a clear solution is obtained.

[0210] Upon addition of 320 mg MgCl2 (4.0 molar equivalents relative to compound 1), the solid metal complex salt immediately began to precipitate.

[0211] The mixture was kept at 20-30°C with stirring for 120 minutes, then 15 mL of diisopropyl ether was added, and the mixture was cooled to 0-10°C and kept with stirring for another 120 minutes.

[0212] Finally, the solid metal complex salt was separated by filtration, washed with 10 mL of diisopropyl ether, dried under nitrogen for 60 minutes, and then dried under vacuum overnight, with the temperature maintained between 0°C and 10°C.

[0213] Yield: 1.30g solid metal complex salt.

[0214] Example 18 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 5.0 g of compound 1 in 50 mL of ethyl acetate and stir until a clear solution is obtained.

[0215] 440 mg of MgCl2 (1.1 molar equivalents relative to compound 1) was added to 10 mL of ethyl acetate. Then, the ethyl acetate solution of compound 1 was added in three portions according to the following steps: first, 25% of the volume of the solution was added at 25°C and stirred for 10 minutes; then, another 25% of the volume of the solution was added at 25°C and stirred for 10 minutes; finally, the remaining 50% of the volume of the solution was added at 25°C and stirred for another 30 minutes. During the addition of the ethyl acetate solution of compound 1, a metal complex salt began to form.

[0216] Then add 75 mL of diisopropyl ether within 1 hour, and keep the mixture at 25 °C with stirring for 180 minutes.

[0217] Finally, the solid metal complex salt was separated by filtration, washed with two equal portions of diisopropyl ether (2 × 10 mL), and dried under nitrogen for 2 hours.

[0218] 4.75 g of solid metal complex salt was obtained and stored at -20 °C.

[0219] Example 19 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) Dissolve 5.0 g of compound 1 in 50 mL of ethyl acetate and stir until a clear solution is obtained.

[0220] 480 mg of MgCl2 (1.2 molar equivalents relative to compound 1) was added to 10 mL of ethyl acetate. Then, the ethyl acetate solution of compound 1 was added in three portions according to the following steps: first, 25% of the volume of the solution was added at 25°C and stirred for 10 minutes; then, another 25% of the volume of the solution was added at 25°C and stirred for 10 minutes; finally, the remaining 50% of the volume of the solution was added at 25°C and stirred for another 30 minutes. During the addition of the ethyl acetate solution of compound 1, a metal complex salt began to form.

[0221] Then add 75 mL of diisopropyl ether within 1 hour, and keep the mixture at 25 °C with stirring for 180 minutes.

[0222] Finally, the solid metal complex salt was separated by filtration, washed with two equal portions of diisopropyl ether (2 × 10 mL), and dried under nitrogen for 2 hours.

[0223] 5.10 g of solid metal complex salt was obtained and stored at -20 °C.

[0224] Example 20 - Synthesis and isolation of magnesium (II) solid complex (solvent: dimethyl carbonate) Dissolve 5.0 g of compound 1 in 25 mL of dimethyl carbonate and stir until a clear solution is obtained.

[0225] 440 mg of MgCl2 (1.0 molar equivalent relative to compound 1) was added to 25 mL of dimethyl carbonate. Then, an ethyl acetate solution of compound 1 was added in three portions according to the following steps: first, 25% of the solution was added at 25°C and stirred for 10 minutes; then, another 25% of the solution was added at 25°C and stirred for 10 minutes; finally, the remaining 50% of the solution was added at 25°C and stirred for another 30 minutes. During the addition of the dimethyl carbonate solution of compound 1, a metal complex salt began to form.

[0226] Then add 150 mL of diisopropyl ether over 1.5 hours and keep the mixture at 25°C with stirring for 180 minutes.

[0227] Finally, the solid metal complex salt was separated by filtration, washed with two equal portions of diisopropyl ether (2 × 10 mL), and dried under nitrogen for 2 hours.

[0228] Example 21 - Synthesis of compound of formula 2 - attachment of side chains of tirzepatide tBuOCO-(CH2) 18 A solution of -CO-Glu(AEEA-AEEA)-OtBu)-OH (3.0 g, 3.43 mmol), OxymaPure® (0.73 g, 5.15 mmol), and DIC (0.531 mL, 3.43 mmol) in DCM was stirred for 2 hours. Then, 0.100 mL of AcOH was added, followed by Fmoc-Lys-OH (2.48 g, 5.15 mmol). The pH was adjusted to 8 with DIPEA, and the reaction mixture was stirred overnight. After the reaction was complete (HPLC controlled), the organic solvent was evaporated. The suspension was diluted with 50 mL of 10% NaHSO4, and the product was extracted twice with 30 mL of ethyl acetate. The organic phase was washed twice with 100 mL of 10% NaHSO4 and 100 mL of water, dried over MgSO4, and then evaporated. The product was purified by flash chromatography and eluted with CHCl3 / methanol mixtures of 20:1, 15:1, 10:1, and 7:1 (v / v) to give 2.31 g of Fmoc-Lys(tBuOCO-(CH2)). 18 -CO-Glu(AEEA-AEEA)-OtBu)-OH (yield 55%, HPLC purity greater than 95%).

[0229] Example 22 - Synthesis and isolation of magnesium (II) solid complex (solvent: ethyl acetate) A suspension of MgCl2 (0.019 g, 0.205 mmol) in 1.1 mL of ethyl acetate was stirred at room temperature, and then Fmoc-Lys(tBuOCO-(CH2)) was added in three equal portions every 30 minutes. 18 A solution of -CO-Glu(AEEA-AEEA)-OtBu)-OH (0.23 g, 0.186 mmol) in 1.1 mL of ethyl acetate was prepared. Then, 3.3 mL of diisopropyl ether was added, and the reaction mixture was stirred for 4 hours. The precipitate was separated by centrifugation, washed twice with 1 mL of diisopropyl ether, and dried under vacuum overnight (yield: 0.24 g, purity 95%).

[0230] Example 23 - Synthesis of tirzepatide fragment 21-29 Peptide fragments 21-29 were synthesized via a stepwise SPPS method using 2-chlorotriphenylmethyl chloride resin (250 mg, moderate loading 1.6 mmol / g). After the resin swelled in 5 mL of DCM, a DCM solution of Fmoc-Gly-OH and DIPEA (0.8 eq and 3 eq, respectively, depending on the resin loading) was added. The reaction mixture was stirred for 1 hour, and unreacted sites on the resin were capped for 30 minutes with a 5 mL mixture of DCM / DIPEA / MeOH (v / v / v 17 / 2 / 1). The resin was then treated with 5 mL of DCM / DIPEA / Ac2O solution and washed with DMF (3 × 3 mL). The resin loading was examined by UV adsorption measurement of the deprotected Fmoc solution and found to be 1.24 mmol / g.

[0231] Then, Fmoc-Ala-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, and Fmoc-Ala-OH (at 3 equivalents relative to resin loading) were pre-activated at room temperature with DIC (3 equivalents) and OxymaPure® (3 equivalents) for 5 minutes, and then continuously coupled to the resin over 60 minutes. Double coupling was performed in the cases of Gln, Val, and Phe. Deprotection of the intermediate Fmoc was performed by washing the resin with DMF (5 × 3 mL) using a 20% piperidine DMF solution (2 × 2.5 mL, for 5 minutes and 20 minutes, respectively).

[0232] Example 24 - Synthesis of tirzepatide fragment 20-29 - attachment of side chains Using 1.5 equivalents of Fmoc-Lys(tBuOCO-(CH2)) 18 The side chain was introduced into the peptide sequence using -CO-Glu(AEEA-AEEA)-OtBu)-OH (in free or complexed form, as described in Example 22), preactivated with 1.5 equivalents of DIC and 1.5 equivalents of OxymaPure®, and then coupled at 40°C for 6 hours. After synthesis, Fmoc protection was removed using a 20% piperidine DMF solution (2 × 2.5 mL, for 5 min and 20 min, respectively). The resin was then washed with DMF (3 × 3 mL) and DCM (2 × 3 mL) and dried. Cleavage was performed in a mixture of TFA / TIS / water (95 / 2.5 / 2.5 v / v / v) to check the coupling efficiency. After cleavage, unprotected fragments 20-29 were obtained.

[0233] Figure 5The HPLC chromatograms of the unprotected fragments 20-29 of telpolide were reported (C18 core-shell column (4.6 × 100 mm)). Eluent A: TFA / H₂O 0.1% v / v; Eluent B: TFA / MeCN 0.1% v / v; Detection wavelength: 220 nm; Gradient elution: 0 min - 5% eluent B; 3 min - 5% eluent B; 33 min - 95% eluent B; 38 min - 95% eluent B; 40 min - 5% eluent B; 50 min - 5% eluent B).

[0234] Specifically, the upper figure refers to fragment 20-29 obtained by coupling the free (i.e., uncomplexed) side chain with fragment 21-29, while the lower figure refers to unprotected fragment 20-29 obtained by coupling the side chain, which has been complexed by reacting with MgCl2 metal salt, with fragment 21-29.

[0235] It can be clearly seen that, with the use of complexed side chains, coupling with fragments 20-29 was also successfully achieved.

[0236] Example 25 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone) Dissolve 20.0 g (16.71 mmol) of compound 1 in 120 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0237] 1.67 mg (17.55 mmol) of anhydrous MgCl2 (1.05 molar equivalent relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 60 minutes. After the addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0238] Then add 360 mL of diisopropyl ether at 20-25°C. Cool the mixture to 10-15°C and maintain it at 10-15°C under nitrogen atmosphere with stirring for 3-4 hours.

[0239] Finally, the solid metal complex salt was separated by filtration, washed with diisopropyl ether (40 mL), and dried at 15-20 °C for 15 hours.

[0240] 19.6 g of solid metal complex salt was obtained and stored at -20 °C.

[0241] Example 26 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, buffer: ammonium acetate) Dissolve 5.0 g (4.18 mmol) of compound 1 in 30 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0242] 100 mg (1.29 mmol) of ammonium acetate (0.3 molar equivalents relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C with stirring for 20 minutes under nitrogen atmosphere. Then, 440 mg (4.60 mmol) of anhydrous MgCl2 (1.10 molar equivalents relative to compound 1) was added, and the mixture was kept at 20–25 °C with stirring for 75 minutes under nitrogen atmosphere. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0243] Then add 90 mL of diisopropyl ether at 20-25 °C. Cool the mixture to 10-15 °C, maintain it at 10-15 °C under nitrogen atmosphere with stirring for 2.5 hours, and then dry it at 10-15 °C for 12 hours.

[0244] 5.5 g of solid metal complex salt was obtained and stored at -20 °C.

[0245] Example 27 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, buffer: sodium ascorbate) Dissolve 1.0 g (0.836 mmol) of compound 1 in 6 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0246] Under nitrogen atmosphere, 80 mg (0.418 mmol) of sodium ascorbate (0.5 molar equivalents relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C with stirring for 20 minutes. Then, 80 mg of anhydrous MgCl2 (1.05 molar equivalents relative to compound 1) was added, and the mixture was kept at 20–25 °C with stirring for 75 minutes. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0247] Then add 18 mL of diisopropyl ether at 20-25°C. Cool the mixture to 10-15°C and maintain it at 10-15°C under nitrogen atmosphere with stirring for 3-4 hours.

[0248] Finally, the solid metal complex salt was separated by filtration, washed with diisopropyl ether (2 mL), and dried at 15-20°C for 15 hours.

[0249] 1.0 g of solid metal complex salt was obtained and stored at -20 °C.

[0250] Example 28 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, weak base: triethylamine) Dissolve 2.0 g (1.67 mmol) of compound 1 in 12 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0251] 170 mg (1.67 mmol) of triethylamine (1.0 molar equivalent relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 10 minutes. Then, 167 mg (1.755 mmol) of anhydrous MgCl2 (1.05 molar equivalent relative to compound 1) was added, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 60 minutes. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0252] Then add 36 mL of diisopropyl ether at 20-25°C. Cool the mixture to 10-15°C and maintain it at 10-15°C under nitrogen atmosphere with stirring for 3-4 hours.

[0253] Finally, the solid metal complex salt was separated by filtration, washed with diisopropyl ether (4 mL), and dried at 15-20°C for 15 hours.

[0254] 2.1 g of solid metal complex salt was obtained and stored at -20 °C.

[0255] Example 29 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, weak base: triethylamine) Dissolve 2.0 g (1.67 mmol) of compound 1 in 12 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0256] 190 mg (1.839 mmol) of triethylamine (1.1 molar equivalents relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 10 minutes. Then, 167 mg (1.755 mmol) of anhydrous MgCl2 (1.05 molar equivalents relative to compound 1) was added, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 60 minutes. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0257] Then add 36 mL of diisopropyl ether at 20-25°C. Cool the mixture to 10-15°C and maintain it at 10-15°C under nitrogen atmosphere with stirring for 3-4 hours.

[0258] Finally, the solid metal complex salt was separated by filtration, washed with diisopropyl ether (4 mL), and dried at 15-20°C for 15 hours.

[0259] 2.1 g of solid metal complex salt was obtained and stored at -20 °C.

[0260] Example 30 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, weak base: triethylamine) 5.2 g (4.35 mmol) of compound 1 was dissolved in 31.2 mL of acetone and stirred at 20-25 °C until a clear solution was obtained.

[0261] 90 mg (0.869 mmol) of triethylamine (0.2 molar equivalents relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 10 minutes. Then, 430 mg of anhydrous MgCl2 (1.05 molar equivalents relative to compound 1) was added, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 60 minutes. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0262] Then add 93.5 mL of diisopropyl ether at 20-25°C. Cool the mixture to 10-15°C and maintain it at 10-15°C under nitrogen atmosphere with stirring for 3-4 hours.

[0263] Finally, the solid metal complex salt was separated by filtration, washed with diisopropyl ether (10 mL), and dried at 15-20°C for 15 hours.

[0264] 5.2 g of solid metal complex salt was obtained and stored at -20 °C.

[0265] Example 31 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, buffer: K-Oxyma) Dissolve 5.0 g (4.18 mmol) of compound 1 in 30 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0266] 791 mg (4.39 mmol) of K-Oxyma (1.05 molar equivalent relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C with stirring for 20 minutes under nitrogen atmosphere. Then, 420 mg (4.39 mmol) of anhydrous MgCl2 (1.05 molar equivalent relative to compound 1) was added, and the mixture was kept at 20–25 °C with stirring for 75 minutes under nitrogen atmosphere. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0267] Then add 90 mL of diisopropyl ether at 20-25 °C. Cool the mixture to 10-15 °C, maintain it at 10-15 °C under nitrogen atmosphere with stirring for 2.5 hours, and dry it under vacuum (<10 Mbar) at 10-15 °C for 12 hours.

[0268] 5.6 g of metal complex salt was obtained and stored at -20 °C.

[0269] Example 32 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, buffer: sodium tetraborate) Dissolve 5.0 g (4.18 mmol) of compound 1 in 30 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0270] 883 mg (4.39 mmol) of sodium tetraborate (1.05 molar equivalent relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 20 minutes. Then, 420 mg (4.39 mmol) of anhydrous MgCl2 (1.05 molar equivalent relative to compound 1) was added, and the mixture was kept at 20–25 °C under nitrogen atmosphere with stirring for 75 minutes. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0271] Then add 90 mL of diisopropyl ether at 20-25 °C. Cool the mixture to 10-15 °C, maintain it at 10-15 °C under nitrogen atmosphere with stirring for 2.5 hours, and then vacuum dry at 10-15 °C for 12 hours.

[0272] 5.8 g of metal complex salt was obtained and stored at -20 °C.

[0273] Example 33 - Synthesis and isolation of magnesium (II) solid complex (solvent: acetone, buffer: potassium tetraborate) Dissolve 5.0 g (4.18 mmol) of compound 1 in 30 mL of acetone and stir at 20-25 °C until a clear solution is obtained.

[0274] 1025 mg (4.39 mmol) of potassium tetraborate (1.05 molar equivalent relative to compound 1) was added to an acetone solution of compound 1, and the mixture was kept at 20–25 °C with stirring for 20 minutes under nitrogen atmosphere. Then, 420 mg (4.39 mmol) of anhydrous MgCl2 (1.05 molar equivalent relative to compound 1) was added, and the mixture was kept at 20–25 °C with stirring for 75 minutes under nitrogen atmosphere. Upon addition of MgCl2 to the acetone solution of compound 1, a gel-like metal complex salt began to form.

[0275] Then add 90 mL of diisopropyl ether at 20-25 °C. Cool the mixture to 10-15 °C, maintain it at 10-15 °C under nitrogen atmosphere with stirring for 2.5 hours, and then vacuum dry at 10-15 °C for 12 hours.

[0276] 5.9 g of metal complex salt was obtained and stored at -20 °C.

[0277] Example 34 - Characterization of metal complex salts Using 1D and 2D in CDCl3 1 The structure of the metal complex salt was studied by H-NMR experiments.

[0278] 1 H-NMR is the application of nuclear magnetic resonance to NMR spectroscopy, focusing on the hydrogen-1 nucleus within a molecule to determine its molecular structure. The proton NMR spectra of most organic compounds exhibit chemical shifts ranging from +14 ppm to -4 ppm and characteristic spin-spin coupling between protons. The integral curve for each proton reflects its abundance.

[0279] Three different samples were analyzed: compound of formula 1 (F1) and metal complex salts obtained by reacting compound of formula 1 with 1 molar equivalent of MgCl2 (C1) and 2 molar equivalents of MgCl2 (C2), respectively.

[0280] The sample 1 H-NMR spectra are shown in Figure 6 In particular, F1 1 H-NMR spectra are shown in Figure 6 a) C1 1 H-NMR spectra are shown in Figure 6 b), C2 1 H-NMR spectra are shown in Figure 6 c). The figure highlights the hydrogen signal positions of the NH group belonging to Glu, the two AEEA groups, and the α-NH and ε-NH groups of Lys.

[0281] Experimental results show that the addition of magnesium chloride reduces the number of amide protons. 1 The significant change in the 1H-NMR signal position indicates that coordination may have occurred between the nitrogen atom of the amine (NH) group and the metal cation. Furthermore, this rules out the possibility of metal salt formation and confirms that the structure obtained from the reaction of compound 1 with MgCl2 is a metal complex salt. If a salt were formed and the amount of metal cation changed, a proton shift would not be expected.

[0282] Here, the resulting positional changes clearly depend on the amount of metal salt added.

[0283] To investigate this dependency in greater depth, multiple two-dimensional studies were conducted on the samples. The CYANA program was used to analyze the samples in the absence and presence of MgCl2. 1 Two-dimensional nuclear overhauser effect (NOESY) spectroscopy in C2 showed a significant increase in the number of intra- and inter-residual nuclear overhauser effects (NOEs) compared to F1 and C1 (Table 1). CYANA ( 1 The combined attribution and kinetic algorithm for H-NMR applications is a type of algorithm based on 1 A program for automatically calculating the structure of biological macromolecules using H-NMR conformation constraints.

[0284] Table 1. NOESY spectral analysis of F1, C1 and C2

[0285] The results of chemical shift perturbation (CSP) analysis further confirmed this trend, confirming that protons 14, 16, and 17 in the AEEA region and the tert-butyl group of glutamic acid appear to be most sensitive to salt addition. This chemical shift perturbation was used to calculate the theoretical spatial arrangement of compounds of formula 1. According to this analysis, compounds of formula 1 have different spatial arrangements depending on the presence and amount of MgCl2 used, such as... Figure 7 As shown, these are compounds of formula 1 ( Figure 7 a) The compound of formula 1 obtained after reacting with 1 molar equivalent of MgCl2 ( Figure 7 b) and the compound of formula 1 after reacting with 2 molar equivalents of MgCl2 ( Figure 7 c).

[0286] Abbreviations

Claims

1. A solid metal complex salt, obtained by reacting a compound of formula I with an MX metal salt: R1-(AEEA)m-A1-CO-(CH2)n-COO-R2 (I), in The amount of the MX metal salt used is in the range of 0.5 to 5.0 molar equivalents relative to the compound of formula I; M is a metal cation selected from alkaline earth metal cations and transition metal cations; X is an organic or inorganic anion; R1 is selected from OH, protected or unprotected amino acids, and peptide groups consisting of 2 to 4 protected or unprotected amino acids; R2 is an H or carboxyl protecting group; A1 is a protected or unprotected amino acid, or a peptide group consisting of 2 to 4 protected or unprotected amino acids; AEEA is 2-[2-(2-aminoethoxy)ethoxy]acetyl; m is an integer between 1 and 10; and n is an integer between 8 and 28.

2. The solid metal complex salt according to claim 1, wherein M is selected from the group consisting of Ca, Zn, Mg, Cu, Co, Ni, Ti, Zr, Fe and Ag cations.

3. The solid metal complex salt according to claim 1 or claim 2, wherein X is a halide anion, preferably X is Cl. - or Br - .

4. The solid metal complex salt according to any one of claims 1 to 3, wherein the amount of the MX metal salt is in the range of 0.8 to 4.0 molar equivalents relative to the compound of formula I, preferably in the range of 1.0 to 2.5 molar equivalents.

5. The solid metal complex salt according to any one of claims 1 to 4, wherein R1 is protected or unprotected Lys and / or Al is protected or unprotected Glu.

6. The solid metal complex salt according to any one of claims 1 to 5, wherein m is an integer between 1 and 5.

7. The solid metal complex salt according to any one of claims 1 to 6, wherein n is an integer between 10 and 25.

8. The solid metal complex salt according to any one of claims 1 to 7, wherein the compound of formula I is a compound of formula 1: （1） Or compounds of formula 2: （2） Or compounds of formula 3: （3） Or compounds of formula 4: （4）。 9. A method for preparing a solid metal complex salt according to any one of claims 1 to 8, comprising the following steps: (i) Disperse or dissolve the compound of formula I in a first equal part of a liquid medium; (ii) Treating compound I with an MX metal salt to form a metal complex salt, and (iii) Recover metal complex salts in solid form.

10. The method for preparing a solid metal complex salt according to claim 9, wherein step (iii) includes antisolvent precipitation or removal of the liquid medium, preferably step (iii) includes antisolvent precipitation.

11. The method for preparing a solid metal complex salt according to claim 9 or claim 10, wherein the liquid medium is selected from the group consisting of water, methanol, isopropanol, ethanol, dichloromethane, chloroform, ethyl acetate, methyl acetate, isopropyl acetate, acetonitrile, dimethyl carbonate, toluene, tetrahydrofuran, acetone, and mixtures thereof.

12. The method for preparing a solid metal complex salt according to any one of claims 9 to 11, wherein the solid metal complex salt is recovered by precipitation with an antisolvent, the antisolvent being selected from the group consisting of heptane, toluene, acetonitrile, ethyl acetate, cyclopentylmethyl ether, petroleum ether, diisopropyl ether, methyl tert-butyl ether, acetone, and mixtures thereof.

13. The method for preparing a solid metal complex salt according to any one of claims 9 to 12, wherein the MX metal salt is first dissolved or dispersed in a second portion of a liquid medium before treating the compound of formula I with the MX metal salt.

14. The method for preparing a solid metal complex salt according to any one of claims 10 to 13, wherein the solid metal complex salt is recovered by antisolvent precipitation, the liquid medium is acetone, ethyl acetate or dimethyl carbonate, and the antisolvent is acetone, ethyl acetate or isopropyl ether, or a mixture of acetone and isopropyl ether, or a mixture of ethyl acetate and isopropyl ether.

15. Use of the solid metal complex salt of any one of claims 1 to 8 in the synthesis of smegglutinin, telposide, and pharmaceutically acceptable salts thereof.

16. A method for preparing an active pharmaceutical ingredient, said active pharmaceutical ingredient comprising smegglutinin, telpoxetine, and pharmaceutically acceptable salts thereof, characterized in that... Use the solid metal complex salt according to any one of claims 1 to 8.

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