Methods for synthesizing targeting molecules

Abstract

The present application provides methods for synthesizing targeting molecules of Formula (I) comprising a retinoid moiety useful for synthesizing liposoluble compounds for targeting and enhancing the activity of therapeutic molecules, including siRNA.

Description

Technical Field

[0001] This application provides a method for synthesizing a targeting molecule comprising a retinoid moiety that can be used to synthesize a lipid-soluble compound for targeting and enhancing the activity of a therapeutic molecule, the therapeutic molecule comprising siRNA. Background Technology

[0002] It is known in the art that generating molecules capable of targeting specific receptors, tissue types, or target organs is highly advantageous. However, when specificity is low, efficacy is limited, requiring larger doses of the therapeutic agent, and off-target effects increase. Molecular scaffolds capable of targeting the delivery of therapeutic agents are one way to overcome these problems. U.S. Patent No. 9,393,315 discloses such a scaffold comprising a basic structure (targeting portion). j -Connector- (Targeting Section) k The targeting portion is a retinol-like group, and j and k are independently 0, 1, 2 or 3; and the linker is PEG-like.

[0003] This scaffold is used to facilitate drug delivery to target cells with specific receptors or activation / binding for retinoids. An example of a molecule using this scaffold is N... 1 N 19 -Bis((S,23E,25E,27E,29E)-16-((2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-en-1-yl)non-2,4,6,8-tetraenamido)-24,28-dimethyl-15,22-dioxo-30-(2,6,6-trimethylcyclohex-1-en-1-yl)-4,7,10-trioxa-14,21-diazatriacontane-23,25,27,29-tetraen-1-yl)-4,7,10,13,16-pentaoxanonadecanediamide (“DiVa”). Previous synthetic strategies for producing DiVa and similar compounds have been hampered by low yields, numerous byproducts, and a lack of readily available high-quality starting materials.

[0004] There is still a need for a synthetic method to produce these retinoid-containing target molecules that promotes increased product yields, low byproduct yields, and the use of more readily available starting materials. Summary of the Invention

[0005] In one aspect, this application provides a method for producing DiVa.

[0006] In one embodiment, this application provides a method for synthesizing compounds of formula I.

[0007]

[0008] Where R 1 It is a retinol-like group;

[0009] m is an integer from 1 to 6; and

[0010] n is an integer between 1 and 10.

[0011] The method includes

[0012] a) Make the compound of formula II

[0013]

[0014] Where m is an integer from 1 to 6;

[0015] Compounds of Formula III

[0016]

[0017] Among them PG 1 and PG 2 Each is an independent protecting group;

[0018] The reaction forms a compound of formula IV.

[0019]

[0020] Where m is an integer from 1 to 6; and

[0021] PG 1 It is a protecting group

[0022] b) Mixing the compound of formula IV with the compound of formula V

[0023]

[0024] Where n is an integer from 1 to 10;

[0025] The reaction forms a compound of formula VI.

[0026]

[0027] Where m is an integer from 1 to 6;

[0028] n is an integer from 1 to 10; and

[0029] PG 1 It is a protecting group;

[0030] c) Reacting the compound of formula VI under hydrogenation conditions to form the compound of formula VII.

[0031]

[0032] Where m is an integer from 1 to 6; and

[0033] n is an integer from 1 to 10; and

[0034] d) React the compound of formula VII with retinoids under coupling conditions to form the compound of formula I.

[0035] In yet another embodiment, the retinoid is selected from vitamin A, retinoic acid, retinoic acid, adapalene, 4-hydroxy(phenyl)retinamide, retinyl palmitate, retinaldehyde, saturated retinoic acid, all-trans retinoic acid, and saturated demethylated retinoic acid.

[0036] In yet another embodiment, the retinoid is retinoic acid.

[0037] In yet another embodiment, the hydrogenation conditions of step c) include reacting the compound of formula VI with H2 and Pd / C.

[0038] In another implementation, m is 3.

[0039] In yet another implementation, n is 5.

[0040] In another implementation, PG 1 and PG 2 Each of the following groups is independently selected from carboxybenzyl, p-methoxybenzylcarbonyl, tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, acetyl, trifluoroacetyl, benzoyl, benzyl, carbamate, p-methoxybenzyl, 3,4-dimethoxybenzyl, p-methoxybenzyl, toluenesulfonyl, trichloroethyl chloroformate, (4-nitrophenyl)sulfonyl, methyl, ethyl, propyl, n-butyl, tert-butyl, succinimide, 2,6-dimethylphenol, 2,6-diisopropylphenol, 2,6-ditert-butylphenol, trimethylsilyl, allyl, 1,1-dimethylallyl, 2,2,2-trifluoroethyl, phenyl, and 4-methoxybenzyl.

[0041] In yet another implementation scheme, each PG 1 It is carboxybenzyl.

[0042] In another implementation, PG 2 It is succinimide.

[0043] In yet another embodiment, the compound of formula II is

[0044] .

[0045] In another embodiment, the compound of formula III is

[0046] .

[0047] In yet another implementation, step a) is performed between -20ºC and 0ºC.

[0048] In yet another implementation, the yield of step a) is at least about 60%. In yet another implementation, the yield of step b) is at least about 70%.

[0049] In yet another embodiment, in step a), the compound of formula II is used in amounts between about 5 and about 20 equivalents, and the compound of formula III is used in amounts of about 1 equivalent.

[0050] In another embodiment, step a) further includes adding citric acid after forming the compound of formula IV.

[0051] In yet another embodiment, the compound of formula I is

[0052] .

[0053] In yet another embodiment, the compound of formula I is produced by the compound of formula III in a yield of at least about 50%.

[0054] In yet another embodiment, the compound of formula IV is produced at a ratio greater than about 9:1 to the compound of formula VIII.

[0055]

[0056] In yet another embodiment, the compound of formula IV is produced at a ratio greater than 12:1 to the compound of formula VIII.

[0057] In another embodiment, the compound of formula I is prepared by the method of steps a)-d). Detailed Implementation

[0058] Throughout the specification and appended claims, the given chemical formula or name shall include all its stereo and optical isomers and racemates, where such isomers are present. Unless otherwise indicated, all chiral (enantiomers and diastereomers) and racemic forms are within the scope of this invention. Many geometric isomers of C=C double bonds, C=N double bonds, cyclic systems, etc., may also be present in the compounds, and all such stable isomers are contemplated in this invention. Cis- and trans- (or E- and Z-) geometric isomers of the compounds of this invention are described and can be separated into mixtures of isomers or isolated isomeric forms. The compounds of this invention can be separated in optically active or racemic forms. Optically active forms can be prepared by resolving racemic forms or by synthesis from optically active starting materials. All methods used to prepare the compounds of this invention and the intermediates prepared therein are considered part of this invention. When preparing enantiomers or diastereomers, they can be separated by conventional methods, such as chromatography or fractional crystallization.

[0059] Depending on the method conditions, the end products of this invention can be obtained in free (neutral) or salt form. Both the free form and the salt of these end products are within the scope of this invention. If desired, one form of the compound can be converted to another. A free base or acid can be converted to a salt; a salt can be converted to a free compound or another salt; a mixture of isomers of the compounds of this invention can be separated into individual isomers. The compounds of this invention, their free forms, and salts can exist in a variety of tautomeric forms, wherein hydrogen atoms are transposed to other parts of the molecule, and thus the chemical bonds between the atoms of the molecule are rearranged. It should be understood that all tautomeric forms, wherever they may exist, are included within the scope of this invention.

[0060] The term "stereoisomer" refers to isomers that have the same composition but differ in the spatial arrangement of their atoms. Enantiomers and diastereomers are examples of stereoisomers. The term "enantiomer" refers to one of a pair of molecular species that are mirror images of each other and cannot be superimposed. The term "diastereomer" refers to a stereoisomer that is not a mirror image. The term "racemic mixture" or "racemic mixture" refers to a composition consisting of two enantiomer species in equimolar amounts, wherein the composition is not optically active. It is well understood in the art that the stereochemistry of the product can be controlled by selecting the stereochemistry of the starting material, and that the stereochemistry of the product can be changed by changing the stereochemistry of the starting material. It is also well understood in the art how to separate racemic mixtures such that the stereochemical purity of the product is > 99%.

[0061] The symbols “R” and “S” represent the configuration of the substituents surrounding one or more chiral carbon atoms. The isomer descriptors “R” and “S” are used as described herein to indicate one or more atomic configurations relative to the core molecule and are intended to be used as defined in the literature (IUPAC Recommendations 1996, Pure and Applied Chemistry, 68, 2193-2222 (1996)).

[0062] The term "chirality" refers to a structural feature of a molecule that prevents it from superimposing itself on its mirror image. The term "isochirality" refers to the state of enantiomer purity. The term "optical activity" refers to the degree to which isochiral molecules or non-racemic mixtures of chiral molecules rotate the plane of polarized light.

[0063] The abbreviations used in this article are defined as follows: ºC represents degrees Celsius, eq represents equivalent, g represents gram, mg represents milligram, L represents liter, mL represents milliliter, µL represents microliter, N represents equivalent concentration, M represents molar concentration, mmol represents millimole, min represents minute, h represents hour, rt represents room temperature, RT represents retention time, conc. represents concentrate, sat or saturated represents saturation, MW represents molecular weight, ee represents enantiomer excess, MS or Mass Spec represents mass spectrometry, ESI represents electrospray ionization mass spectrometry, HR represents high resolution, HRMS represents high resolution mass spectrometry, LCMS represents liquid chromatography-mass spectrometry, HPLC represents high performance liquid chromatography, and NMR represents nuclear magnetic resonance spectroscopy. 1 "H" represents the proton, and "D", "L", "α", "β", "R", "S", "E" and "Z" are stereochemical names familiar to those skilled in the art.

[0064] This paper provides a method for synthesizing a basic structure (R) 1 ) j -Connector-(R) 1 ) k Methods for targeting molecules, where R 1 It is a retinol-like substance; j and k are independently 0, 1, 2, or 3; the connector is PEG-like. In one embodiment, the method is a method for synthesizing compounds of formula I. In one embodiment, the method is a method for synthesizing compounds of formula I starting from compounds of formula III.

[0065] Step a)

[0066] In one implementation, step a) includes

[0067] Compounds of Formula II

[0068]

[0069] Where m is an integer from 1 to 10;

[0070] Compounds of Formula III

[0071]

[0072] Among them PG 2 It is a protecting group;

[0073] The reaction forms a compound of formula IV.

[0074]

[0075] In one embodiment, m is an integer from 1 to 10. In another embodiment, m is an integer from 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In one embodiment, m is an integer from 1 to 6. In another embodiment, m is an integer from 2 to 10, 3 to 10, 4 to 10, 5 to 10, 6 to 10, 7 to 10, 8 to 10, or 9 to 10. In another embodiment, m is an integer from 2 to 9, 3 to 8, 4 to 7, or 5 to 6. In one embodiment, m is 1. In one embodiment, m is 2. In one embodiment, m is 3. In one embodiment, m is 4. In one embodiment, m is 5. In one embodiment, m is 6. In one embodiment, m is 7. In one embodiment, m is 8. In one embodiment, m is 9. In one embodiment, m is 10.

[0076] In one implementation, PG 1 It is an amine protecting group. In another embodiment, PG1 is selected from carboxybenzyl (CBz), p-methoxybenzylcarbonyl (Moz), tert-butoxycarbonyl (BOC), 9-fluorenylmethoxycarbonyl (Fmoc), acetyl (Ac), trifluoroacetyl, benzoyl (Bz), benzyl (Bn), carbamate, p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl, p-methoxybenzyl, toluenesulfonyl (Ts), trichloroethyl chloroformate (Troc), and (4-nitrophenyl)sulfonyl (Nosyl). In one embodiment, PG 1 It is carboxybenzyl.

[0077] In one implementation, PG 2 It is a carboxylic acid protecting group. In one embodiment, PG 2Selected from methyl, ethyl, propyl, n-butyl, tert-butyl, Bn, succinimide (Su), 2,6-dimethylphenol, 2,6-diisopropylphenol, 2,6-di-tert-butylphenol, trimethylsilyl, allyl, 1,1-dimethylallyl, 2,2,2-trifluoroethyl, phenyl (Ph), and 4-methoxybenzyl, in one embodiment, PG 2 It is succinimide.

[0078] In one implementation, PG 1 It is carboxybenzyl, and PG 2 It is succinimide. In one embodiment, PG 1 It is carboxybenzyl, PG 2 It is succinimide, and m is 3.

[0079] In one embodiment, step a) is carried out in a solvent. In one embodiment, the solvent is a combination of solvents. In one embodiment, at least one solvent is polar aprotic. In another embodiment, more than one solvent is polar aprotic. In one embodiment, the one or more solvents are selected from dichloromethane (DCM), ethyl acetate (EtOAc), tetrahydrofuran (THF), acetone, N,N-dimethylformamide (DMF), acetonitrile, and dimethyl sulfoxide (DMSO). In one embodiment, the solvent is a combination of DCM and EtOAc.

[0080] In one embodiment, step a) is performed between about -30ºC and about 0ºC. In one embodiment, the process is performed between about -25ºC and about 0ºC, about -20ºC and about 0ºC, about -15ºC and about 0ºC, or about -10ºC and about 0ºC. In one embodiment, step a) is performed between about -30ºC and about -5ºC, about -30ºC and about -15ºC, or about -30ºC and about -25ºC. In one embodiment, step a) is performed between about -25ºC and about -5ºC, about -20ºC and -10ºC, or about -18ºC and -12ºC. In one embodiment, step a) is performed between about -20ºC and about -12ºC. In one embodiment, step a) is performed at about -20ºC, about -15ºC, about -10ºC, about -5ºC, or about 0ºC. In one embodiment, step a) is performed below about 0ºC. In one embodiment, step a) is performed below about -5ºC. In one embodiment, step a) is performed below about -10ºC. In one embodiment, step a) is performed below about -15ºC.

[0081] In one embodiment, the compound of formula IV is produced from step a) in a yield of about 70% to 95%. In another embodiment, the compound of formula IV is produced in a yield of about 70% to 85% or about 70% to 75%. In yet another embodiment, the compound of formula IV is produced in a yield of about 80% to about 95% or about 90% to 95%. In yet another embodiment, the compound of formula IV is produced in a yield of about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In still another embodiment, the compound of formula IV is produced in a yield of about 80%. In one embodiment, the compound of formula IV is produced from step a) in a yield of at least about 65%, at least 70%, at least 75%, at least about 80%, or at least about 85%. In yet another embodiment, formula IV is produced from step a) in a yield of at least about 80%.

[0082] In one embodiment, step a) includes reacting about 10 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III. In one embodiment, step a) includes reacting about 9 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III, about 8 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III, about 7 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III, about 6 equivalents of the compound of formula II with about 1 equivalent of the compound of formula II, or about 5 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III.

[0083] In one embodiment, step a) includes reacting less than about 8 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III. In one embodiment, step a) includes reacting less than about 7 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III. In one embodiment, step a) includes reacting less than about 6 equivalents of the compound of formula II with about 1 equivalent of the compound of formula III.

[0084] In yet another embodiment, the compound of formula IV is produced at a ratio greater than 9:1 to the compound of formula VIII. In one embodiment, the compound of formula IV is produced at a ratio greater than about 8:1, greater than about 9:1, greater than about 10:1, greater than about 11:1, greater than about 12:1, greater than about 13:1, greater than about 14:1, or greater than about 15:1 to the compound of formula VIII. In another embodiment, the compound of formula IV is produced at a ratio of about 9:1 to the compound of formula VIII. In one embodiment, the compound of formula IV is produced at a ratio of about 8:1, greater than about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, or about 15:1 to the compound of formula VIII.

[0085] In one embodiment, step a) is selective monoamidation of the diamine CAS #4246-51-9 with the bis-Z-lysine derivative CAS #21160-83-8. In one embodiment, reacting the compound of formula III with the compound of formula II in step a) comprises slowly adding a solution of formula III to a solution of formula II. In one embodiment, reacting the compound of formula III with the compound of formula II in step a) comprises adding a solution of formula III subsurface to a solution of formula II.

[0086] Step b

[0087] In one embodiment, step b includes reacting the compound of formula IV with the compound of formula V.

[0088]

[0089] Where n is an integer from 1 to 10;

[0090] The reaction forms a compound of formula VI.

[0091]

[0092] Where m is 1-6;

[0093] n is 1-10; and

[0094] PG 1 It is a protecting group.

[0095] In one implementation scheme, m, n, PG 1 and PG 2 As described in step a). In one implementation, PG 1 It is carboxybenzyl, PG 2 It is succinimide, m is 3, and n is 5.

[0096] In one embodiment, step b) is carried out in a solvent. In one embodiment, at least one solvent is a polar aprotic solvent. In one embodiment, the one or more solvents are selected from dichloromethane (DCM), ethyl acetate (EtOAc), tetrahydrofuran (THF), acetone, N,N-dimethylformamide (DMF), acetonitrile, and dimethyl sulfoxide (DMSO). In one embodiment, step b) is carried out in DCM.

[0097] In one embodiment, step b) is performed between about 0°C and about 30°C. In one embodiment, the process is performed between about 10°C and about 30°C or between about 20°C and about 30°C. In one embodiment, step b) is performed between about 0°C and about 25°C, between about 0°C and about 15°C, or between about 0°C and about 5°C. In one embodiment, step b) is performed between about 15°C and about 20°C. In one embodiment, step b) is performed at about 30°C, about 25°C, about 20°C, about 15°C, about 10°C, about 5°C, or about 0°C. In one embodiment, step b) is performed below about 30°C. In one embodiment, step b) is performed below about 25°C. In one embodiment, step b) is performed at about 20°C. In one embodiment, step b) is performed at about room temperature.

[0098] In one embodiment, the compound of formula VI is produced from step b) in a yield between about 70% and 99%. In another embodiment, the compound of formula VI is produced in a yield between about 70% and 95%, about 70% and 90%, about 70% and 85%, about 70% and 80%, or about 70% and 75%. In another embodiment, the compound of formula VI is produced from step b) in a yield between about 75% and about 99%, about 80% and about 99%, about 85% and about 99%, about 90% and about 99%, or about 95% and about 99%. In yet another embodiment, the compound of formula VI is produced from step b) in a yield of about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99%. In yet another embodiment, the compound of formula VI is produced in a yield of about 90%. In yet another embodiment, the compound of formula VI is produced in a yield of about 95%.

[0099] In one embodiment, the compound of formula VI is generated from step b) in a yield of at least about 65%, at least about 70%, at least about 75%, at least about 80%, or at least about 85%. In another embodiment, formula VI is generated from step b) in a yield of at least about 80%.

[0100] In one embodiment, step b) is the diamidation of 3-(((ethylimino)methylene)amino)-N,N-dimethylpropyl-1-amine (EDAC)-mediated diacid CAS: 439114-13-3.

[0101] Step c

[0102] c) Reacting the compound of formula VI under hydrogenation conditions to form the compound of formula VII.

[0103]

[0104] Where m is an integer from 1 to 10; and

[0105] n is an integer from 1 to 10.

[0106] In one implementation, m and n are as described in step a). In one implementation, m is 3 and n is 5.

[0107] In one embodiment, step c) is carried out in a solvent. In one embodiment, the solvent is a combination of solvents. In another embodiment, at least one solvent is polar proton. In another embodiment, at least one solvent is polar aproton. In another embodiment, the one or more solvents are selected from methanol, ethanol, ethyl acetate, cyclohexane, methylcyclohexane, benzene, petroleum ether, petroleum ether, 2-methyltetrahydrofuran, acetone, tetrahydrofuran, dimethylacetamide, N-methylpyrrolidine, and DMF.

[0108] In one embodiment, the hydrogenation conditions include a hydrogenation catalyst. In one embodiment, the hydrogenation catalyst is selected from palladium, rhenium, rhodium, ruthenium, platinum, or Raney nickel. Other typical hydrogenation catalysts that can be applied are outlined in Wang, D. et al., Chem. Rev. 2015, 115, 13, 6621-6686.

[0109] In one embodiment, the hydrogenation catalyst is on a support. In one embodiment, the support is selected from carbon, alumina, alkaline earth carbonates, clay, ceramics, pumice, or diatomaceous earth. In one embodiment, the support is carbon.

[0110] In one embodiment, the hydrogenation catalyst is palladium, and the support is carbon (carbon-supported palladium or Pd / C).

[0111] In one embodiment, the hydrogenation conditions include a hydrogen donor. In one embodiment, the hydrogen donor is selected from 1-methyl-1,4-cyclohexadiene and 1,4-cyclohexadiene. Other applicable hydrogen donors are outlined in Wang, D. et al., Chem. Rev. 2015, 115, 13, 6621-6686.

[0112] It was found that dissolved oxygen halted the hydrogenation reaction due to side reactions with the hydrogen donor. It was also found that CO2 produced during the hydrogenation reaction halted the reaction due to catalyst poisoning. Therefore, in one embodiment, step c) is carried out in a reaction vessel with an inert atmosphere. In another embodiment, the reaction vessel is sprayed with N2 before the reaction. In yet another embodiment, the reaction vessel is sprayed with N2 throughout the entire hydrogenation reaction.

[0113] In one embodiment, step c) is performed between about 30ºC and about 60ºC. In one embodiment, the process is performed between about 40ºC and about 60ºC or between about 50ºC and about 60ºC. In one embodiment, step c) is performed between about 30ºC and about 50ºC or between about 30ºC and about 40ºC. In one embodiment, step c) is performed between about 25ºC and about 55ºC, between about 30ºC and about 50ºC, between about 35ºC and about 45ºC, or between about 38ºC and about 42ºC. In one embodiment, step c) is performed between about 40ºC and about 45ºC. In one embodiment, step c) is performed at about 30ºC, about 35ºC, about 40ºC, about 45ºC, about 50ºC, about 55ºC, or about 60ºC. In one embodiment, step c) is performed below about 60ºC. In one embodiment, step c) is performed at a temperature below about 50ºC. In another embodiment, step c) is performed at about 45ºC.

[0114] In one embodiment, the compound of formula VII is produced from step b) in a yield between about 70% and 99%. In another embodiment, the compound of formula VII is produced in a yield between about 70% and 99%, about 70% and 95%, about 70% and 90%, about 70% and 85%, about 70% and 80%, or about 70% and 75%. In yet another embodiment, the compound of formula IV is produced from step b) in a yield between about 75% and about 99%, about 80% and about 99%, about 85% and about 99%, about 90% and about 99%, or about 95% and about 99%.

[0115] In another embodiment, the compound of formula VII is produced from step b) in a yield of about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99%. In yet another embodiment, the compound of formula VII is produced in a yield of about 90%. In yet another embodiment, the compound of formula VII is produced in a yield of about 95%. In another embodiment, the compound of formula VII is produced in a yield of about 98%. In yet another embodiment, the compound of formula VII is produced in a yield of about 99%.

[0116] In one embodiment, the compound of formula VII is generated from step b) in a yield of at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99%. In another embodiment, formula VII is generated from step b) in a yield of at least about 80%.

[0117] In one embodiment, step c) is a hydrogenation reaction. In another embodiment, step c) is a heterogeneous palladium-catalyzed transfer hydrogenation of a compound of formula VI.

[0118] Step d

[0119] In one embodiment, step d includes reacting the compound of formula VII with R. 1 The reaction forms a compound of formula I.

[0120] In one implementation, m and n are as described in step a). In one implementation, m is 3 and n is 5.

[0121] In yet another implementation scheme, R 1 The retinoid is selected from vitamin A, retinoic acid, retinoic acid, adapalene, 4-hydroxy(phenyl)retinamide, retinyl palmitate, retinal, all-trans retinoic acid, saturated retinoic acid, all-trans retinoic acid, and saturated demethylated retinoic acid. In one embodiment, the retinoid is retinoic acid. In one embodiment, the retinoid is all-trans retinoic acid.

[0122] In one embodiment, the coupling conditions include an activator. In one embodiment, the activator is selected from 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminoonium tetrafluoroborate (TBTU), (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylureonium hexafluorophosphate (HBTU), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylureonium tetrafluoroborate (TATU), and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM).

[0123] In one embodiment, the coupling condition includes a base. In one embodiment, the base is any base with pKb ≥ 8. In one embodiment, the base is selected from trimethylamine, sodium hydroxide, N-methylmorpholine (NMM), 1-methylimidazole (NMI), N,N-diisopropylethylamine (DiPEA), and potassium hydroxide.

[0124] In one embodiment, step d) further includes purifying the compound of formula I by chromatography. In one embodiment, the chromatography is performed on a silica column. In yet another embodiment, the silica column is a C2 silica column. 18 Column. In one embodiment, the silica column is treated with a solvent gradient. In one embodiment, the solvent gradient comprises 20%-40% ethanol in methanol.

[0125] Compounds of Formula I are found to degrade under light of 500 nm or lower. Therefore, in one embodiment, the compound of Formula I in step d) is inaccessible to light of 500 nm or lower. In one embodiment, the coupling conditions include excluding light of 500 nm or lower. In one embodiment, the purification of the compound of Formula I includes excluding light of 500 nm or lower.

[0126] Compounds of Formula I are found to oxidize upon exposure to the atmosphere. Therefore, in one embodiment, the compound of Formula I in step d) is kept away from oxygen. In one embodiment, step d) is carried out in a reaction vessel with an inert environment. In another embodiment, the reaction vessel is sprayed with N2 prior to the reaction. In another embodiment, step d) further includes loading butylated hydroxytoluene (BHT) into the reaction vessel. In another embodiment, step d) further includes exchanging the solvent for ethanol and concentrating to provide a final 35 wt% solution of the compound of Formula I with 400-1500 ppm BHT to remove oxygen.

[0127] Compounds of Formula I have been found to be relatively unstable above 45ºC. Therefore, in one embodiment, step d) is performed between about 25ºC and about 45ºC. In one embodiment, the process is performed between about 35ºC and about 45ºC or between about 40ºC and 45ºC. In one embodiment, step d) is performed between about 25ºC and about 40ºC, between about 25ºC and about 35ºC, or between about 25ºC and about 30ºC. In one embodiment, step d) is performed between about 30ºC and about 40ºC. In one embodiment, step d) is performed between about 25ºC and about 35ºC. In one embodiment, step d) is performed at about 25ºC, about 30ºC, about 35ºC, about 40ºC, or about 45ºC. In one embodiment, step d) is performed below about 45ºC.

[0128] In one embodiment, the compound of Formula I is produced from step d) in a yield between about 70% and 95%. In another embodiment, the compound of Formula I is produced in a yield between about 70% and 90%, about 70% and 80%, or about 70% and 75%. In another embodiment, the compound of Formula I is produced in a yield between about 80% and about 95%, about 85% and about 95%, or about 90% and 95%. In yet another embodiment, the compound of Formula I is produced in a yield of about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. In yet another embodiment, the compound of Formula I is produced in a yield of about 70%. In yet another embodiment, the compound of Formula I is produced in a yield of about 80%. In one embodiment, the compound of Formula I is produced from step d) in a yield of at least about 65%, at least 70%, at least 75%, at least about 80%, or at least about 85%. In another implementation, Formula I is produced from step d) with a yield of at least about 70%.

[0129] In one embodiment, step d) involves coupling the compound of formula VII and retinoic acid in DCM:2-MeTHF and triethylamine using TBTU. In another embodiment, step d) involves coupling the compound of formula VII and retinoic acid in a mixture of DCM:2-MeTHF and triethylamine at about 30°C by activation with TBTU. In yet another embodiment, step d) involves coupling the compound of formula VII and retinoic acid in DCM:2-MeTHF and triethylamine using TBTU. In yet another embodiment, step d) involves coupling the compound of formula VII and all-trans-retinoic acid in DCM:2-MeTHF and triethylamine at 30°C by activation with TBTU.

[0130] Overall process

[0131] In one embodiment, the compound of Formula I is produced from the compound of Formula III in a total yield of about 40% to about 80%. In one embodiment, the compound of Formula I is produced from the compound of Formula III in a yield of about 40% to about 70%, about 40% to about 60%, about 40% to about 50%, or about 40% to about 45%. In one embodiment, the compound of Formula I is produced from the compound of Formula III in a yield of about 50% to about 80%, about 60% to about 80%, about 70% to about 80%, about 75% to about 80%, about 80% to about 80%, or about 85% to about 80%. In one embodiment, the compound of Formula I is produced from the compound of Formula III in a yield of about 60%. In one embodiment, the compound of formula I is generated from the compound of formula II in a yield of at least about 40%, at least about 50%, at least about 60%, or at least about 65%. In one embodiment, the compound of formula I is generated from the compound of formula III in a yield of at least about 50%. In one embodiment, the compound of formula I is generated from the compound of formula III in a yield of at least about 60%.

[0132] Example

[0133] Example 1. Synthesis of dibenzyl (1-amino-15-oxo-4,7,10-trioxa-14-azaeicosano-16,20-diyl)(S)-dicarbamate

[0134]

[0135] 2-MeTHF (1.71 kg, 2.0 L, 2.0 L / kg) was loaded into reactor 1 and stirring was started. 2,5-Dioxopyrrolidone-1-ylN was added at 20ºC. 2 N 6 -Bis((benzyloxy)carbonyl)-L-lysine ester (1.00 kg) was charged into reactor 1. DCM (7.96 kg, 6.0 L, 6.0 L / kg) was charged at 20ºC. Stirring was started. 2-MeTHF (4.27 kg, 5.0 L, 5.0 L / kg) was charged into reactor 1. 2-MeTHF (5.98 kg, 7.0 L, 7.0 L / kg) was charged into reactor 2 and stirring was started. 3,3'-((oxybis(ethane-2,1-diyl))bis(oxy))bis(propane-1-amine) (3.45 kg, 8.0 equivalent, 3.45 kg / kg) was charged into reactor 2. Reactor 2 was cooled to -25ºC to -12ºC (target -20ºC) and stirring was started. 2,5-Dioxopyrrolidine-1-N from reactor 1 was added. 2 N6 The bis((benzyloxy)carbonyl)-L-lysine ester solution was slowly added to the diamine solution in reactor 2 at < -12ºC, with thorough stirring and subsurface addition via a dip tube. 2,5-Dioxopyrrolidine-1-N 2 N 6 The addition of bis((benzyloxy)carbonyl)-L-lysine ester to 3,3'-((oxybis(ethane-2,1-diyl))bis(oxy))bis(propane-1-amine) is exothermic. Maintain batch temperature ≤ -15ºC. Age the batch for at least 5 minutes with continuous stirring. Reduce the stirring speed and warm reactor 2 to 15ºC-25ºC (target 20ºC). Add 25 wt% brine (8.50 kg, 8.50 kg / kg), followed by water (4.80 kg, 4.80 kg / kg). The addition of brine is exothermic. Maintain batch temperature ≤ 25ºC. Stir this batch at 15ºC-25ºC (target 20ºC) for at least 30 minutes. Separate the phases, sending the bottom aqueous phase to waste. Add 4 wt% citric acid (12.5 kg, 12.5 kg / kg) to reactor 2. Stir this batch at 15ºC–25ºC (target 20ºC) for at least 30 minutes. Separate the phases, sending the product-rich bottom aqueous phase to reactor 1. Return the aqueous layer from reactor 1 to reactor 2. Charge 2-MeTHF (11.53 kg, 13.5 L, 13.5 L / kg) into reactor 2, followed by 10 M NaOH (approx. 1.0 kg, approx. 0.75 L, 1.0 kg / kg) to pH 12–13. Separate the phases, sending the bottom aqueous phase to waste. Charge 25 wt% brine (11.8 kg, 10.0 L, 11.8 kg / kg) into reactor 2, followed by water (5.0 kg, 5.0 kg / kg). Separate the phases, sending the bottom aqueous phase to waste. Concentrate the batch to a final volume of 4.5 L (4.5 L / kg) (approx. 20 wt%) by distillation, while maintaining a temperature ≤ 40ºC. (1-Amino-15-oxo-4,7,10-trioxa-14-azaeicosaecan-16,20-diyl)(S)-dicarbamate dibenzyl ester was synthesized in 88% yield.

[0136] Example 2: Synthesis of tetrabenzyl ((5S,57S)-6,22,40,56-tetraoxo-11,14,17,25,28,31,34,37,45,48,51-undecano-7,21,41,55-tetraazahexadecane-1,5,57,61-tetrayl)tetrabenzyl carbamate

[0137]

[0138] (1-Amino-15-oxo-4,7,10-trioxa-14-azaeicosano-16,20-diyl)(S)-dicarbamate dibenzyl ester (2.05 equivalents) was charged into reactor 1 as a process solution (e.g., 18.7 kg, 20 wt%) at 20ºC. DCM (23.9 kg, 18.0 L / kg) was charged into reactor 1 at 20ºC. Stirring was started and the batch was heated to 15ºC–25ºC (target 20ºC). 4,7,10,13,16-pentaenodecanoic acid (1.00 kg) was charged into reactor 1 at 20ºC. Hydroxybenzotriazole hydrate (HOBT) (45.3 g, 0.100 equivalents) was charged into reactor 1 at 20ºC. EDAC (1.47 kg, 2.60 equivalents) was then charged. The addition of EDAC is exothermic. Maintain batch temperature < 30ºC (target 20ºC). Age the batch at 15ºC-25ºC (target 20ºC) for at least 3 hours. J = 2-MeTHF (43.0 kg, 50.0 L, 50.0 L / kg) was added at 20ºC.

[0139] Add 5.00 wt% citric acid solution (31.0 kg, 30.0 L, 30.0 L / kg). Start stirring and heat the batch to 30ºC–40ºC (target 35ºC). Stir the batch at 30ºC–40ºC (target 35ºC) for 30 minutes. Stop stirring and allow each phase to stand for at least 30 minutes. Separate the phases, sending the bottom aqueous phase to waste.

[0140] Add the 5.00 wt% potassium carbonate solution from step 7 (20.9 kg, 20.0 L, 20.0 L / kg). Start stirring and heat the batch to 25ºC–35ºC (target 30ºC). Stir the batch at 25ºC–35ºC (target 30ºC) for 30 minutes. Stop stirring and allow each phase to stand for at least 30 minutes. Separate the phases, sending the bottom aqueous phase to waste.

[0141] Add 25 wt% sodium chloride brine (11.94 kg, 10.00 L, 10.00 L / kg). Add water (11.94 kg, 11.94 L, 11.94 L / kg). Stir the batch at 25ºC–35ºC (target 30ºC) for 30 minutes. Stop stirring and allow each phase to stand for at least 30 minutes. Separate the phases, sending the bottom aqueous phase to the waste.

[0142] Begin stirring and cool the batch to 0ºC. Concentrate the batch to a final volume of 12.5 L (12.5 L / kg), maintaining the batch temperature. 50ºC. For product stability and distillation rate, distillation is recommended at Tj ≤ 50ºC and P ≤ 80 mbar. MeOH (9.9 kg, 12.5 L, 12.5 L / kg) was loaded through a spray nozzle. The solution was discharged at room temperature through a polished filter (pore size ≤ 10 μm) to store the BMT-334119 process solution. Tetrabenzyl tetracarbamate ((5S,57S)-6,22,40,56-tetraoxo-11,14,17,25,28,31,34,37,45,48,51-undecano-7,21,41,55-tetraazahexadecane-1,5,57,61-tetrayl) was produced in 90%–95% yield.

[0143] Example 3: N 1 N 19 Synthesis of bis((S)-16,20-diamino-15-oxo-4,7,10-trioxa-14-azaeicosyl)-4,7,10,13,16-pentaoxa-nonadecanidamide.

[0144]

[0145] Two reactors (R1 and R2) were set up with a transfer line between them. Both reactors were kept at 20ºC. Tetrabenzyl tetracarbamate ((5S,57S)-6,22,40,56-tetraoxo-11,14,17,25,28,31,34,37,45,48,51-undecano-7,21,41,55-tetraazahexahexadecane-1,5,57,61-tetrayl)tetrabenzyl tetracarbamate (1.00 kg) was added to R2 as a process solution in a 1:1 2-MeTHF:MeOH mixture. 1-Methyl-1,4-cyclohexadiene (920 g, 1.10 L, 15.0 equivalents) was then added to R2. MeOH (4.11 kg, 5.2 L, 5.2 L / kg) and 2-MeTHF (0.512 kg, 0.6 L, 0.6 L / kg) were added to R1. N2 was injected into R1 and R2 for ≥ 20 min. Subsurface nitrogen flow was achieved by feeding through an immersion tube near the impeller. 5% Pd / C (100 g, 0.100 kg / kg) was added to R1 under inert conditions. N2 was injected into R1 and R2 for ≥ 20 min.

[0146] Heat the batch in R1 to 40ºC–50ºC (target 45ºC). Transfer the contents of R2 to R1 above the surface. Stir for >3.5 h. At the end of reaction aging, take a sample for reaction completion. Immediately filter the sample (≤1 μm pore size) to quench the reaction. N 1 N 19-Bis((S)-16,20-diamino-15-oxo-4,7,10-trioxa-14-azaeicosyl)-4,7,10,13,16-pentaoxa-nonadecanidamide was produced in 98% yield.

[0147] Example 4: N 1 N 19 Synthesis of bis((S,23E,25E,27E,29E)-16-((2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-en-1-yl)non-2,4,6,8-tetraenamido)-24,28-dimethyl-15,22-dioxo-30-(2,6,6-trimethylcyclohex-1-en-1-yl)-4,7,10-trioxa-14,21-diazatriacontane-23,25,27,29-tetraen-1-yl)-4,7,10,13,16-pentaoxanonadecanediamide

[0148]

[0149] Add water (10 kg, 10 L, 10 l / kg) to reactor 1. Add sodium bicarbonate (0.6 kg, 0.6 kg / kg) to reactor 1. Stir reactor 1 until all solids are visibly dissolved. Add N to a 1:1 mixture of 2-MeTHF:MeOH (1.00 kg). 1 N 19 The process solution of bis((S)-16,20-diamino-15-oxo-4,7,10-trioxa-14-azaeicosyl)-4,7,10,13,16-pentaoxa-nonadecanid was charged into reactor 2. Stirring was started and the batch was cooled to between 5°C and 10°C. N 1 N 19 The solution of bis((S)-16,20-diamino-15-oxo-4,7,10-trioxa-14-azaeicosaedecyl)-4,7,10,13,16-pentaoxa-nonadecanedimamide was concentrated to a final volume of 5 L / kg (5 L) and the solvent was exchanged for 2-MeTHF while maintaining the batch temperature ≤ 45ºC. The batch in reactor 2 was heated to 25ºC–35ºC (target 30ºC). DCM (15 L, 15 L / kg) was charged into reactor 2. All-trans retinoic acid (1.50 kg, 5.00 equivalents) was charged. Triethylamine (1.52 kg, 2.09 L, 15.0 equivalents) was charged. The batch was aged at 25ºC–35ºC (target 30ºC) for at least 30 minutes until all solids were visibly dissolved. TBTU (1.77 kg, 5.5 equivalents) was charged. The batch material shall be aged at 25ºC-35ºC (target 30ºC) for no less than 4 hours.

[0150] Charge BHT (0.0018 kg, 0.0018 kg / kg). Charge 6 wt% sodium bicarbonate into reactor 2 (10.6 kg, 10 L, 10 L / kg). Stir the batch at 25ºC–35ºC (target 30ºC) for at least 1 hour. Stop stirring and allow each phase to stand for at least 30 minutes. Separate the phases in reactor 2, sending the bottom organic phase to reactor 3 and the top aqueous phase to waste. Start stirring in reactor 3 and cool the batch to between 5ºC and 10ºC. Concentrate the batch to 10 L / kg (10 L) and exchange the solvent for MeOH while maintaining the batch temperature ≤ 45ºC. Sample for IPC-4. Charge 200°C (proof) ethanol (0.79 kg, 1 L, 1 L / kg). Use an ethanol:methanol gradient through C 18 Preparative chromatography method for product purification. N 1 N 19 -Bis((S,23E,25E,27E,29E)-16-((2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-en-1-yl)non-2,4,6,8-tetraenamido)-24,28-dimethyl-15,22-dioxo-30-(2,6,6-trimethylcyclohex-1-en-1-yl)-4,7,10-trioxa-14,21-diazatriacontane-23,25,27,29-tetraen-1-yl)-4,7,10,13,16-pentaoxanonadecanediamide was produced in 85% yield.

Claims

1. A method for synthesizing compounds of formula I. Where R 1 It is a retinol-like group; m is an integer from 1 to 6; and n is an integer from 1 to 10; The method includes a) Make the compound of formula II Where m is an integer from 1 to 6; Compounds of Formula III Among them PG 1 and PG 2 Each is an independent protecting group; The reaction forms a compound of formula IV. Where m is an integer from 1 to 6; and PG 1 It is a protecting group b) Mixing the compound of formula IV with the compound of formula V Where n is an integer from 1 to 10; The reaction forms a compound of formula VI. Where m is an integer from 1 to 6; n is 1-10; and PG 1 It is a protecting group; c) Reacting the compound of formula VI under hydrogenation conditions to form the compound of formula VII. Where m is an integer from 1 to 6; and n is an integer from 1 to 10; and d) React the compound of formula VII with retinoids under coupling conditions to form the compound of formula I.

2. The method according to claim 1, wherein the retinoid in step d) is selected from vitamin A, retinoic acid, retinoic acid, adapalene, 4-hydroxy(phenyl)retinamide, retinyl palmitate, retinaldehyde, saturated retinoic acid and saturated demethylated retinoic acid.

3. The method according to claim 2, wherein the retinoid in step d) is retinoic acid.

4. The method according to claim 1, wherein the hydrogenation conditions comprise reacting the compound of formula VI with H2 and Pd / C.

5. The method according to claim 1, wherein m is 3.

6. The method of claim 5, wherein n is 5.

7. The method of claim 1, wherein PG 1 and PG 2 Each of the following groups is independently selected from carboxybenzyl, p-methoxybenzylcarbonyl, tert-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, acetyl, trifluoroacetyl, benzoyl, benzyl, carbamate, p-methoxybenzyl, 3,4-dimethoxybenzyl, toluenesulfonyl, trichloroethyl chloroformate, (4-nitrophenyl)sulfonyl, methyl, ethyl, propyl, n-butyl, tert-butyl, succinimide, 2,6-dimethylphenol, 2,6-diisopropylphenol, 2,6-ditert-butylphenol, trimethylsilyl, allyl, 1,1-dimethylallyl, 2,2,2-trifluoroethyl, phenyl, and 4-methoxybenzyl.

8. The method of claim 7, wherein PG 1 It is carboxybenzyl.

9. The method of claim 7, wherein PG 2 It is succinimide.

10. The method according to claim 1, wherein the compound of formula II is 。 11. The method of claim 10, wherein the compound of formula III is 。 12. The method of claim 1, wherein step a) is performed between -20ºC and 0ºC.

13. The method of claim 1, wherein the yield of step a) is at least 70%.

14. The method of claim 1, wherein in step a), the compound of formula II is present in amounts of 2 to 8 equivalents, and the compound of formula III is present in an amount of 1 equivalent.

15. The method of claim 1, wherein step a) further comprises adding citric acid after forming the compound of formula IV.

16. The method according to claim 1, wherein the compound of formula I is 。 17. The method of claim 1, wherein the compound of formula I is produced from the compound of formula III in a yield of at least 50%.

18. The method of claim 1, wherein the compound of formula IV is produced at a ratio greater than 9:1 to the compound of formula VIII. 。

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