Succinate salts of n-(3-(4-(3-(diisobutylamino) propyl) piperazin-1-yl) propyl)-1h-benzo [d] imidazole-2-amine, their preparation and their use

Abstract

The present invention relates to succinate salts of N-(3-(4-(3-(diisobutylamino) propyl) piperazin-1-yl) propyl)-1H-benzo [d] imidazol-2-amine and pharmaceutically acceptable solvates thereof, their preparation, pharmaceutical compositions containing them, and their use in the treatment and / or prophylaxis of neurodegenerative diseases.

Description

[0001] This application is a divisional application of patent application No. 202180064847.4, filed on September 30, 2021, entitled “Succinate of N-(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1H-benzo[d]imidazol-2-amine, its preparation and use thereof”. Technical Field

[0002] This invention relates to new N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate and its pharmaceutically acceptable solvates, their preparation, pharmaceutical compositions containing them and their use in the treatment and / or prevention of neurodegenerative diseases. Background Technology

[0003] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine, which has the structure of formula I Formula I It is a pharmaceutically active molecule (PAM) that was previously disclosed in WO 2006 / 051489, belongs to the 1,4-bis(3-aminopropyl)piperazine derivative family, and can be used to treat and / or prevent neurodegenerative diseases.

[0004] The free base form of this compound suffers from long-term stability issues due to oxidative degradation. Therefore, the applicant sought a salt form to overcome these stability problems and discovered the salt form described in WO2014 / 102339. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d [Imidazole-2-amine sulfate. This sulfate is not oxidatively degraded and is obtained as a stable powder, exhibiting a crystalline phase with good crystallinity; based on commonly used standards (i.e., water absorption less than 2% by weight at 25°C / 60% relative humidity (RH), it is non-hygroscopic. In particular, the applicant found...] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazol-2-amine disulfate has a shelf life of at least 5 years under normal storage conditions (15-25°C) and exhibits significant water solubility greater than 100 mg / mL. Therefore, N-(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1H-benzo[d]imidazol-2-amine sulfate is a suitable active pharmaceutical ingredient (API).

[0005] Under development N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine disulfate API, used to treat neurodegenerative diseases, has successfully completed a Phase 1 clinical trial in healthy volunteers. This disulfate API was administered orally to healthy volunteers in a liquid formulation: an aqueous solution. These solutions are easy to prepare and very stable (up to 18 months at 5°C, up to 18 months at 25°C / 60% RH, and up to 6 months at 40°C / 75% RH in glass vials) without any preservatives. The same formulation was used in a Phase 2A clinical trial in patients with progressive supranuclear palsy (PSP).

[0006] The applicant hopes to develop N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dSolid dosage forms (e.g., capsules) in the salt form of imidazole-2-amine PAM are preferred, while the free base is excluded due to its instability and lower solubility. In fact, solid dosage forms offer many advantages over liquid dosage forms: easier stability, packaging, supply, storage, use, and logistics, and are more cost-effective. Unlike liquid dosage forms, the development (registration compliance) of solid dosage forms requires the API to be a unique, well-defined polymorphic form. A polymorphic form of a molecule, or polymorph, refers to a different solid state of a molecule that possesses different and unique physicochemical properties, such as crystallinity, melting point, solubility, and stability. The properties of APIs and their solid dosage forms depend on the polymorphic form of the API; polymorphs can exhibit different stability, solubility, and dissolution rates, which can significantly affect the bioavailability of the drug. Therefore, drug approval and regulatory requirements dictate that the solid dosage form of a drug is a unique, stable polymorph of the API; for example, refer to the U.S. Food and Drug Administration's industry guidance on Chemistry, Manufacturing, and Controls (CMC) requirements for submitting Abbreviated New Drug Applications (FDA-2004-D-0182) when a drug substance exists in a polymorphic form. Thus, selecting the correct polymorph of the correct salt is crucial for API development, and employing highly reproducible chemical processes for its synthesis is equally detrimental. In this case, the final step in the API synthesis method is salt formation, which needs to be optimized to directly obtain the selected polymorphic form. In other words, the method will be more efficient and cost-effective when selecting a salt / polymorph form compatible with the final step of one-pot salt formation-direct crystallization.

[0007] The applicant first studied N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine disulfate was subjected to a comprehensive polymorph screening, resulting in the identification of numerous polymorphs. The polymorph named "Type A" has the following XRPD diffraction pattern: Figure 1As shown, the most suitable stable form was identified for the preparation of solid dosage forms. The applicant then sought to develop a reproducible chemical method for the synthesis of type A, namely, identifying suitable conditions (solvent, antisolvent, concentration, duration, temperature, nucleation conditions, filtration, drying conditions) for the one-pot formation of disulfate / direct crystallization from a free base in this polymorphic form. Despite extensive research, the applicant was unable to achieve this result and could only obtain the desired type A polymorph from a free base in two distinct steps: 1) forming disulfate from a polymorph referred to as "type B," and then 2) converting type B to the target type A. The applicant further identified that the target type A was sensitive to drying conditions, as it reverted to polymorph B after drying at 50°C. This sensitivity further limited the industrial-scale approach, and the applicant switched to studying type B, as this polymorph is suitable for the more efficient one-pot salt formation / crystallization final step.

[0008] Type B (its XRPD diffraction pattern is in) Figure 2 (Published in China) was identified from a polymorph screening study. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d It is the most prominent polymorphic form of imidazole-2-amine disulfate. However, there is a risk of crystal form transformation in several different solvents or under high humidity conditions, and it may be prone to salt disproportionation (formation of trisulfate) in alcohol solvents. In addition, form B partially loses its crystallinity after prolonged drying at 50°C. Therefore, the applicant has overlooked form B as a suitable polymorphic form for developing solid dosage forms.

[0009] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine disulfate, despite being a stable, non-hygroscopic crystalline powder API, exhibits complex polymorphism in terms of crystal stability and synthetic process efficiency. Robust and cost-effective synthetic methods employing a one-pot salt formation / direct crystallization final step cannot guarantee the acquisition of a specific stable polymorph. Polymorphism screening criteria are used to determine the appropriate polymorphic form. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The polymorphs of imidazole-2-amine are generally unstable during routine drying at 50°C. The applicant concludes that disulfide is unsuitable for obtaining [the desired product]. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dSolid dosage form of imidazole-2-amine PAM.

[0010] WO2014 / 102339 discussed N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Other salts of imidazole-2-amine. The oxalate initially reported in WO2006 / 051489 was unsuitable due to the potential for nephrotoxicity caused by the oxalate ion. The hydrochloride was found to be highly hygroscopic, while the acetate was not obtained in solid form. The tartrate and fumarate salts obtained were amorphous solids. Malate was discarded because its chirality made clinical development quite complex, and each stereoisomer needed to be equivalently characterized as the active stereoisomer.

[0011] Therefore, it is still necessary in this field N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The stable polymorphic form of the new salt of imidazole-2-amine is particularly suitable for solid dosage forms and can be obtained by stable chemical methods. Summary of the Invention

[0012] This invention is based on an unexpected discovery, namely N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate provides a stable powder; this polymorph exhibits high crystallinity, is very stable, non-hygroscopic, and can be obtained in excellent yield from free base in a one-pot salting / direct crystallization process. Succinate meets the aforementioned requirements and is particularly suitable for use as an API in solid dosage forms, such as capsules.

[0013] Therefore, this invention relates to... N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate and its pharmaceutically acceptable solvates.

[0014] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinates and their solvates are obtained as powders, exhibiting high crystallinity and defined crystalline phases. Furthermore, the succinates of the present invention are particularly suitable for preparing pharmaceutical compositions containing them. The succinates are pharmaceutically acceptable and, to the best of the applicant's knowledge, are not associated with any kind of inherent toxicity.

[0015] Furthermore, compared to other salts such as sulfates, succinates have a simpler polymorphic form with higher crystallinity, are more stable during the synthesis process (especially the conventional 50°C drying) (i.e., no change in crystal form), and do not undergo disproportionation (i.e., stoichiometry) in the solvents used in this method.

[0016] Therefore, the use of succinate will enhance and facilitate the use of substances containing... N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The production of pharmaceutical compositions of imidazole-2-amine PAM, while reducing related costs. Invention Details The compound of the present invention is N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinates and their pharmaceutically acceptable solvates. More particularly, the succinates and solvates of the present invention are those of formula II.

[0018] Formula II Where x is 1-4, preferably x is 1.4-3.1, more preferably x is 1.4-1.6, and even more preferably x is about 1.5, or even more preferably x is 1.5.

[0019] In other words, 1 molecule N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazol-2-amine N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The succinate of imidazole-2-amine contains 1-4 equivalents, preferably 1.4-3.1 equivalents, more preferably 1.4-1.6 equivalents, even more preferably about 1.5 equivalents, and even more preferably 1.5 equivalents of succinate.

[0020] In a preferred embodiment, the succinate of the present invention is N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d ] Sesquisuccinate of imidazole-2-amine.

[0021] In one embodiment, the preferred succinate of the present invention is a compound of formula II, wherein x is 2.9-3.1, preferably wherein x is about 3, and more preferably x is 3.

[0022] In another preferred embodiment, the succinate of the present invention is N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine trisuccinate.

[0023] In one specific embodiment, the succinate of Formula II is a pharmaceutically acceptable solvate, preferably a hydrate. For one molecule of the succinate of Formula II, the solvate stoichiometry is 0.4-2, preferably 0.4-1.2, more preferably 0.5-1.1, even more preferably about 0.5 or about 1.1, and even more preferably 0.5 or 1.1 molecules of solvate.

[0024] In one embodiment, the compound of the present invention is N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate hemihydrate.

[0025] In another embodiment, the compound of the present invention is N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate monohydrate.

[0026] The applicant has identified several polymorphic forms of the compounds of the present invention, which are particularly stable against 1) chemical degradation (such as salt disproportionation) and 2) polymorphic changes. These polymorphs exhibit high crystallinity, are stable under normal storage and stress conditions, and are non-hygroscopic. The applicant has discovered specific experimental conditions for the preparation of these polymorphic forms using the final step of a one-pot salt formation / crystallization process from free base. Therefore, the compounds of the present invention can be used as APIs in solid dosage forms.

[0027] In one embodiment, a particularly preferred compound of the present invention is a compound of formula II, wherein x is 1.5, and whose X-ray powder diffraction (XRPD) pattern, when irradiated with a Cukα light source, contains peaks at diffraction angles of 2θ = 3.8° ± 0.2°, 10.3° ± 0.2°, 12.4° ± 0.2°, 16.2° ± 0.2°, 17.9° ± 0.2°, 19.8° ± 0.2°, 20.4° ± 0.2°, 23.8° ± 0.2°, and 26.7° ± 0.2°. This particular crystal form is referred to as "Crystal Form 1" throughout the application, and its XRPD diffraction pattern is recorded on a PANalytical X'per tpro with a PIXcel detector (128 channels), as shown below. Figure 3 As shown.

[0028] Another preferred compound of the present invention is the compound of formula II, wherein x is 3, and when irradiated with a CuKα light source, its X-ray powder diffraction (XRPD) pattern contains peaks at diffraction angles of 2θ = 5.3° ± 0.2°, 15.0° ± 0.2°, 15.3° ± 0.2°, 16.8° ± 0.2°, 17.9° ± 0.2°, 20.5° ± 0.2°, 21.2° ± 0.2°, 23.9° ± 0.2°, 24.3° ± 0.2°, and 26.6° ± 0.2°. This particular crystal form is referred to as "Crystal Form 2" throughout the application, and its XRPD diffraction pattern is recorded on a PANalytical X'pert pro with a PIXcel detector (128 channels), as shown below. Figure 4 As shown.

[0029] Another preferred compound of the present invention is the compound of formula II, wherein x is 1.5, y is 0.5, and when irradiated with a CuKα light source, its crystal form exhibits an X-ray powder diffraction (XRPD) pattern containing peaks at diffraction angles of 2θ = 3.8° ± 0.2°, 4.4° ± 0.2°, 9.2° ± 0.2°, 10.6° ± 0.2°, 13.8° ± 0.2°, 17.7° ± 0.2°, 19.9° ± 0.2°, 21.6° ± 0.2°, and 23.6° ± 0.2°. Its XRPD diffraction pattern is recorded on a PANalytical X'pert pro with a PIXcel detector (128 channels), as shown below. Figure 5 As shown.

[0030] Another preferred compound of the present invention is the compound of formula II, wherein x is 1.5, y is 1.1, and when irradiated with a CuKα light source, its crystal form exhibits X-ray powder diffraction (XRPD) patterns containing peaks at diffraction angles of 2θ = 4.2 ± 0.2°, 4.4° ± 0.2°, 9.6° ± 0.2°, 10.6° ± 0.2°, 15.0° ± 0.2°, 16.3° ± 0.2°, 17.4° ± 0.2°, 19.7° ± 0.2°, 22.8° ± 0.2°, 29.1°, and 29.7° ± 0.2°. Its XRPD diffraction pattern is recorded in a PANalytical X'pert pro with a PIXcel detector (128 channels), as shown below. Figure 6 As shown.

[0031] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The crystalline form of imidazole-2-amine sesquisuccinate can be obtained by chemical synthesis. This method is another objective of the present invention.

[0032] This invention also relates to the preparation of N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d A method for determining the crystal form of imidazole-2-amine sesquisuccinate, comprising the following steps: - Step 1: Put N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine dissolves in an organic solvent; - Step 2: Heat the reaction medium to temperature B, where temperature B is defined as 30℃-50℃; - Step 3: Add approximately an equimolar amount of succinic acid; - Step 4: Stir the reaction medium at temperature B for 0.25-4 hours; - Step 5: Cycle the reaction medium between temperature A and temperature B for 2-4 hours for 10-30 hours, where temperature A is defined as 0-10℃. - Step 6: Cool the reaction medium to temperature A; - Step 7: Filter the reaction medium at temperature A; - Step 8: Wash the filter cake with the organic solvent at temperature A; - Step 9: Dry the filter cake at a temperature of 30-50℃.

[0033] In the first step, provide N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d A solution of imidazole-2-amine. This can be used to prepare solutions. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The imidazole-2-amine feedstock can be in any physical form of the free base, including solvated forms, with a purity ≥95%. In one embodiment, the organic solvent is selected from the group consisting of acetone, 97.5% acetone / 2.5% water (%v / v), methyl ethyl ketone, acetonitrile, and tert-butyl methyl ether. In a particularly preferred embodiment, the organic solvent used is methyl ethyl ketone, which is particularly suitable for preparing crystal form 1. The volume of the organic solvent used is crucial for the dissolution of the free base and the successful formation and crystallization of its salt. In a preferred embodiment, the amount of organic solvent added is 20-40 mL / g free base, preferably 25-35 mL / g free base, and more preferably about 30 mL / g free base.

[0034] In a preferred embodiment, heating step 2 is performed at a temperature of 35°C-45°C, more preferably 40°C-45°C, even more preferably about 40°C, and even more preferably at a temperature B of 40°C.

[0035] The optional filtration step at temperature B can be performed between step 2 and the subsequent step 3, which involves the addition of succinic acid. Succinic acid is used in relation to... N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The imidazole-2-amine free base is added in substantially an equimolar amount. In other words, 0.95 to 1.05 equivalents (eq.) of succinic acid are added relative to the free base. In a preferred embodiment, 1.0 equivalent of succinic acid is used.

[0036] In step 3, succinic acid is added in batches using powdered succinic acid or by slowly adding a suspension or solution of concentrated succinic acid in an organic solvent used to dissolve the alkali. In a preferred embodiment, succinic acid is added in powdered form. In another preferred embodiment, succinic acid is added as a concentrated solution in an organic solvent.

[0037] In step 4, salt formation begins while the reaction medium is stirred at temperature B for 0.5-4 hours. In a preferred embodiment, the medium is stirred for 20-40 minutes, preferably about 30 minutes. In another preferred embodiment, the medium is stirred for 2-4 hours, preferably about 3 hours.

[0038] The subsequent temperature cycling step 5 causes the sesquisuccinate to crystallize in a specific crystal form. The temperature cycling is performed for 10-30 hours between temperatures A and B, with each cycle lasting 2-4 hours, where temperature A is defined as 0-10°C. In a particularly preferred embodiment, the temperature cycling is performed for about 3 hours between about 5°C and about 40°C, lasting for about 20 hours, thereby causing the sesquisuccinate to crystallize in crystal form 1, especially when methyl ethyl ketone is used as the organic solvent.

[0039] At this point, the crystalline salt is typically precipitated and the reaction medium is cooled to 0-10°C, preferably about 5°C (step 6), and then the desired substance is recovered by filtration at the same temperature (step 7). The filter cake is washed once with the same organic solvent used in the reaction (step 8), and the desired crystalline salt is recovered from the filter cake. In a preferred embodiment, about 5 mL of organic solvent is used per gram of substance during filter cake washing.

[0040] Then, the filtered and washed material is dried, preferably under reduced pressure, at a temperature of 30-50°C, preferably about 40°C (step 9).

[0041] The particularly preferred method of the present invention can obtain N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate type 1, as follows: - Step 1: Put N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine dissolved in methyl ethyl ketone; - Step 2: Heat the reaction medium to 45°C; - Step 3: Add an equimolar amount of succinic acid; - Step 4: Stir the reaction medium at 45°C for 3-4 hours; - Step 5: Cycle the reaction medium at 5℃-40℃ for 3 hours, and then cycle the temperature for 20 hours. - Step 6: Cool the reaction medium to 5°C; - Step 7: Filter the reaction medium at 5°C; - Step 8: Wash the filter cake with the organic solvent at 5°C; - Step 9: Dry the filter cake under reduced pressure at 40°C.

[0042] The method according to the invention makes N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate direct one-pot salt formation / crystallization.

[0043] When the stoichiometry of succinic acid is changed to about 2 equivalents by 1) using ethanol as an organic solvent and 2) changing the stoichiometry of succinic acid to about 2 equivalents in step 3, the method according to the invention makes it possible to N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine trisuccinate direct one-pot salt formation / crystallization.

[0044] The applicant has shown N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinate and its pharmaceutically acceptable solvates can be used to correct the metabolism of Tau protein, particularly by inhibiting pathological Tau protein phosphorylation. Tau protein (also known as microtubule-associated protein Tau-MAPT) interacts with tubulin to stabilize microtubules and promote the assembly of tubulin into microtubules; microtubule stability is controlled by isotype and phosphorylation. Tau pathology includes mechanisms leading to aberrant modifications of microtubule-associated Tau protein, such as its hyperphosphorylation, progressive accumulation, and accumulation in degenerating neurons as fibrillary material to form so-called neurofibrillary tangles (NFTs). The compounds of the present invention are effective in protecting the neuronal synaptic network and promoting neurite growth. Neurons are the processes of the neuronal cell body, and their presence and growth are detrimental to interneuronal connections. Furthermore, the compounds of the present invention are able to reduce neuroinflammation and microglial activation while increasing the level of granulin precursor (PGRN) neurotrophic factor. Neuroinflammation is a secondary response in chronic, progressive neurodegenerative diseases, including but not limited to tau diseases and Parkinson's disease, which can be triggered by activated microglia releasing neurotoxic pro-inflammatory cytokines that cause neuronal loss. PGRN is a secreted glycoprotein primarily expressed in mature neurons and microglia, which maintains the vitality of the cerebral cortex (Brain J. Neurol. 140, 3081-3104 (2017)). PGRN expression in the brain is low during early development and increases with age (Trends Neurosci. 37, 388-398 (2014), J. Reprod. Dev. 57, 113–119 (2011)). However, in response to injury, activated microglia upregulate PGRN expression (Acta Neuropathol. (Berl.) 119, 123-133 (2010), Neuroscience 250, 8-19 (2013)). Regarding Tau pathology, PGRN deficiency accelerates Tau deposition and phosphorylation in mice expressing human tau (J. Neuropathol. Exp. Neurol. 74, 158-165 (2015)). PGRN can be processed into granular proteins by several proteases, including elastase released from activated microglia (Neurology 90, 118-125 (2018)). Human and rodent data have shown that reduced PGRN is associated with increased Tau pathology. PGRN haploinsufficiency increases Tau phosphorylation in P301L-Tau transgenic mice, strongly supporting the idea that reduced PGRN protein may lead to Tau hyperphosphorylation and intraneuronal accumulation (J. Neuropathol. Exp. Neurol. 74, 158-165 (2015)).Furthermore, inflammatory stimulation in the brain has been reported to be associated with increased Tau phosphorylation (J. Neuroinflammation 7, 56 (2010)). Since PGRN can act as an anti-inflammatory factor in neuroinflammation (Neuroscience 250, 8-19 (2013)), it is hypothesized that Tau phosphorylation will be accelerated in inflammatory states induced by reduced PGRN levels. PGRN is a neurotrophic factor whose role in Parkinson's disease is well-studied. Van Kampen et al. have used a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of PD to investigate the potential use of PGRN gene delivery as a therapy for the prevention or treatment of PD (PLOS one 2014, 9, 5, e97032). Viral vector delivery of the PGRN gene is an effective means of increasing PGRN expression in substantia nigra-striatal neurons. When SN... C Elevated PGRN expression in mice protected substantia nigra striatal neurons from MPTP toxicity, while also preserving striatal dopamine levels and turnover. Furthermore, PGRN gene therapy protected substantia nigra striatal neurons with a reduction in MPTP-induced markers of inflammation and apoptosis, as well as complete preservation of motor function. In addition, blood levels of granulin precursors in humans can reflect the severity and progression of Parkinson's disease (Neuroscience Letters, volume 725, 23 April 2020, 134873). Compared to controls, Parkinson's disease patients had significantly lower plasma levels of granulin precursors, which were also found to be negatively correlated with motor symptoms, disease severity, and duration, thus indicating a neuroprotective role for PGRN in PD. All these findings support that increasing PGRN levels could be an effective therapy for improving neurodegenerative diseases such as tau dysplasia and Parkinson's disease. In particular, the succinate of the present invention has been found to have neuroprotective effects and can significantly reduce the effects of mitochondrial toxin MPP. + Accumulation of α-synuclein in dopaminergic neurons in rotenone-damaged dopaminergic neurons (two well-known in vitro models of PD).

[0045] Tau protein diseases include, but are not limited to, Alzheimer's disease, amyotrophic lateral sclerosis (ALS) and Parkinson-Guam dementia syndrome, auricolic granulation disease, chronic traumatic encephalopathy, corticobasal degeneration, pulsatile dementia, diffuse neurofibrillary tangles with calcification, familial British dementia, familial Danish dementia, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granule protein subtype (FTD-GRN), Gerstmann-Sträussler-Scheinker disease, Guadeloupean Parkinson's disease, and Hallewarden-Schpatz disease. Diseases including inclusion body myositis, multiple system atrophy, Steinert's myotonic dystrophy, type II myotonic dystrophy, neurodegeneration with iron buildup in the brain, Niemann-Pick disease type C, Non-Guamanian motor neuron disease with neurofibrillary tangles, Paget's disease, Pick's disease, post-encephalitis Parkinson's syndrome, progressive subcortical glial proliferation, progressive supranuclear palsy (PSP), SLC9A6-related intellectual disability, subacute sclerosing panencephalitis, tangled dementia, multi-infarct dementia, ischemic stroke, chronic traumatic encephalopathy (CTE), traumatic brain injury (TBI), stroke, and white matter tau proteinosis with globular glial inclusions.

[0046] Furthermore, the physicochemical properties (such as crystallinity, stability, and solubility) of the succinates and their solvates of the present invention are particularly useful for solid and liquid pharmaceutical formulations.

[0047] Therefore, as described above, due to its ability to correct Tau protein metabolism, increase PGRN neurotrophic factor levels and reduce neuroinflammation, the succinate and its solvates of the present invention can be used as medicines, particularly for the treatment or prevention of neurodegenerative diseases, including tau protein diseases and Parkinson's disease.

[0048] Therefore, the succinate of the present invention can be used as a medicine, particularly for the treatment or prevention of neurodegenerative diseases and all diseases in which dysfunction of Tau protein phosphorylation is observed, including but not limited to tau protein diseases.

[0049] In one embodiment, the present invention relates to succinates for the treatment or prevention of diseases selected from neurodegenerative diseases and diseases in which impaired phosphorylation of Tau protein is observed, including but not limited to tau protein diseases.

[0050] Therefore, the present invention also relates to the invention as defined herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof for the treatment and / or prevention of diseases selected from: Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson-Guam dementia syndrome, aerobatic granulation disease, chronic traumatic encephalopathy, corticobasal degeneration, muscular dementia, diffuse neurofibrillary tangles with calcification, familial British dementia, familial Danish dementia, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granulin subtype (FTD-GRN), Gerstmann-Sträussler-Scheinker disease, Guadeloup E.G. Parkinson's disease, Hallewarden-Scholes disease, inclusion body myositis, multiple system atrophy, Steinert's myotonic dystrophy, type II myotonic dystrophy, neurodegeneration with brain iron accumulation, Niemann-Pick disease type C, Non-Guamanian motor neuron disease with neurofibrillary tangles, Paget's disease, Pick's disease, post-encephalitis Parkinson's syndrome, progressive subcortical glial proliferation, progressive supranuclear palsy (PSP), SLC9A6-related intellectual disability, subacute sclerosing panencephalitis, tangled dementia, multi-infarct dementia, ischemic stroke, chronic traumatic encephalopathy (CTE), traumatic brain injury (TBI), stroke, and white matter tau proteinosis with globular glial inclusions. Preferably, the disease is selected from Alzheimer's disease, auricula-jugular disease, corticobasal degeneration, diffuse neurofibrillary tangles with calcification, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granular protein subtype (FTD-GRN), Pick's disease, progressive subcortical gliosis, progressive supranuclear palsy (PSP), tangles dementia, and white matter tau proteinopathy with globular glial inclusions. More preferably, the disease is selected from Alzheimer's disease, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granular protein subtype (FTD-GRN), progressive supranuclear palsy (PSP), tangles dementia, and white matter tau proteinopathy with globular glial inclusions. Even more preferably, the disease is selected from Alzheimer's disease and progressive supranuclear palsy (PSP).

[0051] In other respects, the present invention also provides methods for treating and / or preventing diseases selected from tau protein diseases, particularly those tau protein diseases cited above, and embodiments thereof, said methods comprising administering to a patient in need a pharmaceutically effective amount as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1H -benzo[ d [Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof. In one specific embodiment, the disease is selected from Alzheimer's disease and progressive supranuclear palsy (PSP).]

[0052] In other respects, the present invention also provides as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The use of imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof in the preparation of medicaments for the treatment and / or prevention of tau protein diseases, particularly those cited above. In one specific embodiment, the diseases are selected from Alzheimer's disease and progressive supranuclear palsy (PSP).

[0053] In another embodiment, the invention also relates to the succinate of the invention for delaying the onset of diseases selected from neurodegenerative diseases and diseases in which dysfunction of Tau protein phosphorylation is observed, including but not limited to tau protein diseases.

[0054] In one specific embodiment, the invention also relates to what is defined herein as... N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof is used in patients to delay the onset of diseases selected from: Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson-Guam dementia syndrome, aerobatic granulation disease, chronic traumatic encephalopathy, corticobasal degeneration, muscular dementia, diffuse neurofibrillary tangles with calcification, familial British dementia, familial Danish dementia, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granulin subtype (FTD-GRN), Gerstmann-Sträussler-Scheinker disease, Guadeloupe disease. Pean Parkinson's disease, Hallewarden-Scholes disease, inclusion body myositis, multiple system atrophy, Steinert myotonic dystrophy, type II myotonic dystrophy, neurodegeneration with iron buildup in the brain, Niemann-Pick disease type C, non-Guamanian motor neuron disease with neurofibrillary tangles, Paget's disease, Pick's disease, post-encephalitis Parkinson's syndrome, progressive subcortical glial proliferation, progressive supranuclear palsy (PSP), SLC9A6-related intellectual disability, subacute sclerosing panencephalitis, tangled dementia, multi-infarct dementia, ischemic stroke, chronic traumatic encephalopathy (CTE), traumatic brain injury (TBI), stroke, and white matter tau proteinosis with globular glial inclusions. Preferably, the disease is selected from Alzheimer's disease, auricula-jugular disease, corticobasal degeneration, diffuse neurofibrillary tangles with calcification, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal-17 linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granular protein subtype (FTD-GRN), Pick's disease, progressive subcortical gliosis, progressive supranuclear palsy (PSP), tangles dementia, and white matter tau proteinopathy with globular glial inclusions. More preferably, the disease is selected from Alzheimer's disease, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal-17 linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granular protein subtype (FTD-GRN), progressive supranuclear palsy (PSP), tangles dementia, and white matter tau proteinopathy with globular glial inclusions. Even more preferably, the disease is selected from Alzheimer's disease and progressive supranuclear palsy (PSP).

[0055] In other respects, the present invention provides a method and embodiments thereof for delaying the onset of disease in patients selected from tau protein diseases, particularly the tau protein diseases cited above, said method comprising administering to a patient in need a pharmaceutically effective amount of the succinate of the present invention or a pharmaceutically acceptable solvate thereof. In one specific embodiment, said disease is selected from Alzheimer's disease and progressive supranuclear palsy (PSP).

[0056] In other respects, the present invention also provides as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The use of imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof in the preparation of a medicament for delaying the onset of tau protein diseases, particularly those mentioned above, in patients, and embodiments thereof. In one specific embodiment, the diseases are selected from Alzheimer's disease and progressive supranuclear palsy (PSP).

[0057] This invention also relates to, as defined herein, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof, used for the treatment and / or prevention of Parkinson's disease.

[0058] In other respects, the present invention also provides a method for treating and / or preventing Parkinson's disease, comprising administering to a patient in need a pharmaceutically effective amount as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof.

[0059] In other respects, the present invention also provides as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Use of imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof in the preparation of medicaments for the treatment and / or prevention of Parkinson's disease.

[0060] In one specific embodiment, the invention also relates to the invention as defined herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof, used to delay the onset of Parkinson's disease in patients.

[0061] In other respects, the present invention provides a method for delaying the onset of Parkinson's disease in a patient, comprising administering to a patient in need a pharmaceutically effective amount of the succinate of the present invention or a pharmaceutically acceptable solvate thereof.

[0062] In other respects, the present invention also provides as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Use of succinate of imidazole-2-amine or a pharmaceutically acceptable solvate thereof in the preparation of a medicament for delaying the onset of Parkinson's disease in patients.

[0063] According to another feature of the invention, a method is provided for inhibiting pathological Tau protein phosphorylation in patients requiring such treatment, preferably warm-blooded animals, and even more preferably humans, said method comprising administering an effective amount of the invention to said patient. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof.

[0064] According to another feature of the invention, a method is provided for inhibiting pathological Tau protein phosphorylation while increasing the level of neurotrophic factor granule protein precursor (PGRN) in patients requiring such treatment, preferably warm-blooded animals, and even more preferably humans, comprising administering an effective amount of the present invention to the patient. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof.

[0065] The present invention also provides a method comprising as described herein. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d A pharmaceutical composition comprising a succinate of imidazole-2-amine or a pharmaceutically acceptable solvate thereof and at least one pharmaceutically acceptable carrier, diluent, excipient, and / or adjuvant. In one embodiment, the invention also covers pharmaceutical compositions which, in addition to containing N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1H -benzo[ d In addition to the succinate of imidazole-2-amine or its pharmaceutically acceptable solvate as the active ingredient, it also contains other therapeutic agents and / or active ingredients.

[0066] According to one embodiment, the present invention N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinates and their pharmaceutically acceptable solvates can be administered as part of a combination therapy. Therefore, embodiments involving the co-administration of compositions and drugs are included within the scope of this invention, wherein the compositions and drugs contain, in addition to the succinates of this invention or their pharmaceutically acceptable solvates as active ingredients, other therapeutic agents and / or active ingredients. Such multi-drug therapy regimens, commonly referred to as "combination therapy," can be used to treat and / or prevent any neurodegenerative disease, particularly tau disease or Parkinson's disease. The use of such combinations of therapeutic agents is particularly suitable for the treatment of patients who require treatment or are at risk of developing the aforementioned neurodegenerative diseases.

[0067] Apart from the need for therapeutic efficacy (which may require the use of...) N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d In addition to the active agent other than the succinate of imidazole-2-amine or its pharmaceutically acceptable solvate, there may be other reasons compelling or strongly recommending the use of a pharmaceutical combination involving an active ingredient representing adjunctive therapy; that is, its supplementation and enhancement of the active ingredients of the present invention. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The function of imidazole-2-amine succinate or its pharmaceutically acceptable solvates. Suitable adjuvant agents for adjuvant therapeutic purposes include those that, instead of directly treating and / or preventing diseases or conditions mediated by or associated with pathological protein Tau phosphorylation and / or neuroinflammation, treat diseases or conditions directly arising from or indirectly associated with diseases or conditions regulated by underlying or potential pathological protein Tau phosphorylation and / or neuroinflammation.

[0068] According to another feature of the invention, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine succinate, a pharmaceutically acceptable solvate, can be used in combination with other drugs used to treat neurodegenerative diseases such as Alzheimer's disease, progressive supranuclear palsy (PSP), and Parkinson's disease. More specifically, compounds of Formula II and their pharmaceutically acceptable solvates may be used in combination with acetylcholinesterase inhibitors as adjunctive therapy, including but not limited to donepezil (CAS n° 120014-06-4) and its salts and solvates, galantamine (CAS n° 357-70-0) and its salts and solvates, rivastigmine (CAS n° 123441-03-2) and its salts and solvates, tacrine (CAS n° 321-64-2) and its salts and solvates, or in combination with NMDA glutamate antagonists, including but not limited to ammonium nitrate (CAS n° 19982-08-2) and its salts and solvates, or in combination with dual acetylcholinesterase inhibitors and NMDA glutamate antagonists, including but not limited to huperzine A (CAS n° 102518-79-6) and its salts and solvates, or in combination with glucagon-like peptide-1 (GLP-1) agonists, including but not limited to liraglutide (CAS n° 102518-79-6) and its salts and solvates, or in combination with glucagon-like peptide-1 (GLP-1) agonists, including but not limited to liraglutide (CAS n° 102514-06-4). Exenatide (CAS n° 141732-76-5) and its salts and solvates, or in combination with retinoids, including but not limited to acitretin (CAS n° 55079-83-9) and its salts and solvates, or in combination with calcium channel blockers (CCBs), including but not limited to nivadipine (CAS n° 75530-68-6) and its salts and solvates, nifedipine (CAS n° 39562-70-4) and its salts and solvates, nimodipine (CAS n° 66085-59-4) and its salts and solvates, or in combination with angiotensin receptor blockers, including but not limited to valsartan (CAS n° 137862-53-4) and its salts and solvates, or in combination with tetracycline antibiotics, including but not limited to minocycline (CAS n° 204656-20-2). 10118-90-8) and its salts and solvates, or in combination with monoclonal antibodies selected from the group consisting of: aducanumab, bapineuzumab, solanezumab, gantenerumab, crenezumab, BAN2401, GSK933776, AAB-003, SAR228810, BIIB037 / BART, ABBV-8E12, BIIB092, and UCB0107, or in combination with glycosidase (O-GlcNAcase) inhibitors, or in combination with dopamine prodrugs, including but not limited to levodopa, optionally in combination with carbidopa, or in combination with dopamine receptor agonists, including but not limited to pramipexole, rotigotine, ropinirole, and amantadine.It may be combined with anticholinergic drugs, such as benzalkonium chloride and trihexyphenidyl; or with MAO-B inhibitors, including but not limited to safinamide, selegiline, and rasagiline; or with COMT inhibitors, including but not limited to entacapone, octopcapone, and tocapone.

[0069] Therefore, the treatment methods and pharmaceutical compositions of the present invention can be used as a single therapy. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d [Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof. However, the methods and compositions described can also be used for a variety of treatments, wherein...] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d One or more succinates of imidazole-2-amine or pharmaceutically acceptable solvates thereof may be administered in combination with one or more other therapeutic agents.

[0070] In the above implementation scheme, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or its pharmaceutically acceptable solvates, in combination with other therapeutic agents, may be administered in separate or combined doses, and at the time of their administration, sequentially or simultaneously. Thus, the administration of one component agent may be before, simultaneously with, or after the administration of other component agents.

[0071] Generally, for drug use, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate or a pharmaceutically acceptable solvate thereof may be formulated into a pharmaceutical composition comprising at least one succinate or a pharmaceutically acceptable solvate thereof of the present invention and at least one pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant, and optionally one or more other therapeutic agents and / or active ingredients.

[0072] By way of non-limiting example, pharmaceutical compositions may be dosage forms suitable for oral administration, parenteral administration (such as by intravenous, intramuscular, or subcutaneous injection or intravenous infusion), topical administration, inhalation, administration via skin patches, implantation, suppositories, etc. Such suitable forms of administration (which may be solid, semi-solid, or liquid, depending on the method of administration) and the methods and carriers, diluents, and excipients used in their preparation will be clear to those skilled in the art; refer to the latest edition of Remington's Pharmaceutical Sciences. Pharmaceutical compositions may be formulated into solid forms and re-dissolved or suspended prior to use. Preferred pharmaceutical compositions of the succinates or solvates of the present invention are in solid dosage forms suitable for oral administration.

[0073] Some preferred but non-limiting examples of dosage forms include tablets, pills, powders, lozenges, capsules, elixirs, suspensions, emulsions, solutions, syrups, aerosols, ointments, creams, lotions, soft and hard gelatin capsules, suppositories, drops, sterile injectable solutions, and sterile packaged powders (which are typically reconstituted before use) for bolus and / or continuous administration, which may be formulated with carriers, excipients, and diluents that are themselves suitable for such formulations, such as lactose, glucose, sucrose, sorbitol, mannitol, starch, agar, gum arabic, calcium phosphate, alginate, astragalus gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethylene glycol, cellulose, (sterilized) water, methylcellulose, methyl and propylparabens, talc, magnesium stearate, edible oils, vegetable oils, and mineral oils, or suitable mixtures thereof. Particularly preferred dosage forms of the succinates and their solvates of the present invention are soft and hard capsules, particularly soft and hard gelatin capsules, preferably hard gelatin capsules, formulated with at least one microcrystalline cellulose excipient, especially Avicel® PH101. The pharmaceutical composition may optionally contain other substances commonly used in pharmaceutical formulations, such as lubricants, wetting agents, emulsifiers and suspending agents, dispersants, disintegrants, stabilizers, isotonic agents, swelling agents, fillers, preservatives, sweeteners, flavoring agents, aroma agents, coloring agents, antibacterial and / or antifungal agents, such as benzoates, chlorobutanol, phenol, sorbic acid, dispersants, flow conditioners, releasing agents, etc. Compositions can also be formulated to provide rapid, sustained, or delayed release of the active compound contained therein.

[0074] The pharmaceutical compositions of the present invention are preferably in unit dose form and may be suitably packaged, for example in boxes, blister packs, vials, bottles, pouches, ampoules, or any other suitable single- or multi-dose holders or containers (which may be suitably labeled); optionally, having one or more leaflets containing product information and / or instructions for use. Typically, such unit doses will contain 0.05-1000 mg, usually 1-500 mg, of at least one compound of the present invention, for example, about 10, 25, 50, 100, 200, 300, or 400 mg per unit dose.

[0075] Typically, depending on the condition to be prevented or treated and the route of administration, the active compounds of the present invention will be administered daily at a dose of 0.01-100 mg / kg, more typically 0.1-50 mg / kg, such as 1-25 mg / kg, for example about 0.5, 1, 2, 5, 10, 15, 20 or 25 mg / kg of patient weight. This may be administered as a single daily dose, divided into one or more daily doses, or administered substantially continuously, for example by infusion.

[0076] All compounds of formula II mentioned include the solvates mentioned, particularly hydrates, multicomponent complexes, and their liquid crystals.

[0077] The compounds disclosed in full in this application were used with ChemDraw. ® Ultra version 11.0 (Cambridgesoft, Cambridge, MA, USA) named.

[0078] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The free base of imidazole-2-amine can be obtained as disclosed in WO2006 / 051489. Succinates and their solvates can be prepared according to techniques known in the art, such as those involving precipitation, crystallization, recrystallization, freeze-drying, phase transfer, or ion exchange resins.

[0079] definition The following definitions and interpretations apply to the entire application, including terms used in the specification, drawings, and claims.

[0080] When describing the compounds of the present invention, unless otherwise specified, the terminology used shall be interpreted in accordance with the following definitions.

[0081] Unless otherwise stated, any reference to compounds of the present invention herein refers to compounds of formula II and their pharmaceutically acceptable solvates. All references to compounds of formula II include the pharmaceutically acceptable solvates mentioned herein and all their crystalline forms.

[0082] The term "administration" or its variations (e.g., "administering") refers to the application of an active agent or active ingredient (e.g., ...). N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine may be given alone or as part of a pharmaceutically acceptable composition to a patient for the treatment or prevention of their condition, symptoms or disease.

[0083] The term “Alzheimer’s disease” as used in this article refers to all types of Alzheimer’s disease, including but not limited to sporadic and familial types.

[0084] The term "inhibition" as used in this article refers to partial inhibition or reduction, as well as complete inhibition.

[0085] The term "person" refers to both sexes and subjects at any developmental stage (i.e., newborn, infant, adolescent, teenager, adult).

[0086] The term "patient" refers to a warm-blooded animal, more preferably a human, that is awaiting or receiving medical care or is / will become the subject of a medical procedure.

[0087] "Pharmaceutical acceptable" means that the components of a drug composition are compatible with each other and are harmless to the patient.

[0088] As used in this article, “prevent / preventing / prevention” refers to methods that delay or eliminate the onset of a symptom or disease and / or its accompanying symptoms, prevent a patient from acquiring the symptom or disease, or reduce the risk of a patient acquiring the symptom or disease.

[0089] The term "sesquisuccinate" as used in this article refers to 1.5 equivalents, and the sesquisuccinate of a compound refers to its succinic acid addition salt, wherein succinic acid exists in a 3:2 ratio with its free base form.

[0090] The term "solvent" is used herein to describe compounds of the present invention that contain stoichiometric or substoichiometric amounts of one or more pharmaceutically acceptable solvent molecules, such as ethanol. When the solvent is water, the term "hydrate" is used. Pharmaceutically acceptable solvent molecules may co-crystallize with the compounds of the present invention, and / or exist in their solid crystalline and / or amorphous phases, and / or be adsorbed onto them.

[0091] The compounds of the present invention include compounds of formula II as defined above, including all polymorphs and crystalline inertia thereof, their prodrugs, and isotopically labeled compounds of formula II.

[0092] As used herein, the term "Tau disease" refers to a condition characterized by abnormal metabolism of the Tau protein, particularly abnormal phosphorylation and / or hyperphosphorylation of the Tau protein and / or elevated levels (above normal control levels, e.g., above normal control levels in individuals or populations of individuals of the same age) of Tau protein or its pathological forms in cells, tissues, or fluids (preferably brain tissue and / or cerebrospinal fluid). More specifically, Tau disease refers to a condition in which intracellular accumulations of abnormally modified Tau protein are observed, such as neurofibrillary tangles (NFTs), pick bodies, astrocyte clusters, and / or muscle inclusions, preferably at least neurofibrillary tangles (NFTs).

[0093] As used herein, the term "therapeutic effective amount" (or more simply, "effective amount") refers to an active agent or active ingredient (e.g., N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d The amount of imidazole-2-amine is sufficient to achieve the desired therapeutic or preventative effect in the patients to whom it is administered.

[0094] The term “treatment” as used in this article is intended to include the reduction, weakening or elimination of symptoms or diseases and / or their accompanying symptoms.

[0095] The invention will be better understood by referring to the following embodiments. These embodiments are intended to represent specific implementations of the invention and are not intended to limit the scope of the invention. Attached Figure Description

[0096] Figure 1 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of imidazole-2-amine disulfate in its A-type crystalline form.

[0097] Figure 2 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of imidazole-2-amine disulfide in type B crystal form.

[0098] Figure 3 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1H -benzo[ d XRPD 2θ diffraction pattern of the crystal form of imidazole-2-amine sesquisuccinate 1.

[0099] Figure 4 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of the crystal form of imidazole-2-amine trisuccinate.

[0100] Figure 5 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of the crystalline form of imidazole-2-amine sesquisuccinate hemihydrate (0.5 eq. water).

[0101] Figure 6 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of the crystalline form of imidazole-2-amine sesquisuccinate monohydrate (1.1 eq. water).

[0102] Figure 7 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d HPLC chromatogram of the crystal form of imidazole-2-amine sesquisuccinate 1.

[0103] Figure 8 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate: TG / DT thermal analysis diagram of the crystalline form.

[0104] Figure 9 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate: DSC thermal analysis diagram of the crystalline form (first heating cycle).

[0105] Figure 10 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate: GVS isotherm diagram of the crystalline form.

[0106] Figure 11 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate: GVS kinetic diagram of the crystalline form.

[0107] Figure 12 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate. GVS post-XRPD 2θ diffraction pattern of the crystalline form.

[0108] Figure 13 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d FT-IR spectrum of crystal form 1 of imidazole-2-amine sesquisuccinate.

[0109] Figure 14 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate 1 H-NMR spectrum (DMSO-d6, 500.12MHz).

[0110] Figure 15 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 1 of imidazole-2-amine sesquisuccinate 13 C quantitative NMR spectrum (DMSO-d6, 500.12 MHz).

[0111] Figure 16 Displaying 7-day stability test results N-(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD 2θ diffraction pattern of the crystal form of imidazole-2-amine sesquisuccinate 1.

[0112] Figure 17 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Overlapping of XRPD 2θ diffraction patterns of a batch of crystal forms of imidazole-2-amine sesquisuccinate.

[0113] Figure 18 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d ] Crystal form 2 of imidazole-2-amine trisuccinate TG / DT thermal analysis diagram of the crystalline form.

[0114] Figure 19 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d DSC thermal analysis diagram of the crystal form of imidazole-2-amine trisuccinate.

[0115] Figure 20 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystal form 2 of imidazole-2-amine trisuccinate 1 H-NMR spectrum (MeOH-d4, 500.12MHz).

[0116] Figure 21 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d DVS isotherm plot of crystal form 2 of imidazole-2-amine trisuccinate.

[0117] Figure 22 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dDVS kinetic diagram of crystal form 2 of imidazole-2-amine trisuccinate.

[0118] Figure 23 Showing N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD (4-35° 2θ) diffraction pattern of a blend of imidazole-2-amine sesquisuccinate crystal form 1 and Avicel® PH101 formulated in hard gelatin capsules and stored at room temperature.

[0119] Figure 24 show N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d XRPD (4-35° 2θ) diffraction pattern of a blend of imidazole-2-amine sesquisuccinate crystal form 1 and Avicel® PH101 formulated in hard gelatin capsules and stored at 40°C / 75% RH.

[0120] Figure 25 Show Aβ 1-42 72 hours after the injury, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Effects of imidazole-2-amine sesquisuccinate crystal form 1 co-cultured with microglia on survival (A), synaptic network (B), and Tau phosphorylation (AT100) (C) in cortical neurons. Results are expressed as a percentage of the control (CTR) condition, and are presented as mean ± SEM (n=4–6). Fisher's test was performed after one-way ANOVA. ; or This indicates p<0.05; p<0.01 or p<0.001 vs Aβ 1-42 Or, for ### vs CTR, p<0.05 is considered significant.

[0121] Figure 26 Display Aβ 1-42 72 hours after the injury, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dEffects of imidazole-2-amine sesquisuccinate crystal form 1 co-cultured with microglia on microglia activation (A) and PGRN release (B) in cortical neurons. Results are expressed as a percentage of the control (CTR) condition, and are presented as mean ± SEM (n=4–6). Fisher's test was performed after one-way ANOVA. ; or This indicates p<0.05; p<0.01 or p<0.001 vs Aβ 1-42 Or, for ### vs CTR, p<0.05 is considered significant.

[0122] Figure 27 Displayed in MPP + After injury N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Effects of imidazole-2-amine sesquisuccinate crystal form 1 on survival (A), neural synaptic network (B), and α-synuclein accumulation (C) of primary midbrain neurons TH(+) pre-incubated or co-incubated for 48 h. Results are expressed as a percentage of control conditions, and are presented as mean ± SEM (n=4–6). Fisher's test was performed after one-way ANOVA. ####p<0.0001 vs control; <0.05; p<0.01; p<0.001 or p<0.0001 vs MPP + .

[0123] Figure 28 This indicates that rotenone damage occurred 24 hours after the injury. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Effects of pre-incubation or co-incubation of imidazole-2-amine sesquisuccinate crystal form 1 on survival (A), synaptic network (B), and α-synuclein accumulation (C) in primary midbrain neurons TH(+). Results are expressed as a percentage of control and are presented as mean ± SEM (n=4–6). Fisher's test was performed after one-way ANOVA. ####p<0.0001 vs. control; <0.05; p<0.01; p<0.001 or p<0.0001 vs rotenone. Detailed Implementation

[0124] Chemical Examples The following abbreviations are used throughout this application: ℃: degrees Celsius, a w Water activity, DMSO: dimethyl sulfoxide, DSC: differential scanning calorimetry, GVS: gravimetric vapor adsorption, δ: NMR chemical shift (in ppm), eq: equivalent, EDTA: ethylenediaminetetraacetic acid, eq.: equivalent, g: gram, h: hour, HDPE: high-density polyethylene, HPLC: high-performance liquid chromatography, Hz: hertz, FT-IR: Fourier transform infrared spectroscopy, L: liter, M: mol / L, mM: mmol / L, μM: μmol / L, Me: methyl, MEK: methyl ethyl ketone, mg: milligram, MHz: megahertz, min: minute, mL: milliliter, mm: millimeter, mol: mole, mmol: millimole, μmol: micromole, MPP + : 1-Methyl-4-phenylpyridin-1-onium, nm: nanometer, NMR: nuclear magnetic resonance, PBS: phosphate-buffered saline, PLM: polarized light microscopy, PGRN: particulate protein precursor, ppm: parts per million, RH: relative humidity, RPM: revolutions per minute, rt: retention time, RT: room temperature (ca 15-25℃), s: second (s), TFA: trifluoroacetic acid, TG / DTA: thermogravimetric / differential thermal analysis, UV: ultraviolet light, v: volume, VH-XRPD: variable humidity X-ray powder diffraction, WB: Western blot, XRPD: X-ray powder diffraction.

[0125] All listed temperatures are in degrees Celsius (°C); unless otherwise stated, all reactions were carried out at room temperature (RT).

[0126] The experimental setups or purification procedures used in this invention, unless otherwise described in detail, are assumed to be known to those skilled in the art and described in such standard reference manuals as: i) Gordon, AJ; Ford, RA, “The Chemist's Companion - A Handbook of Practical Data, Techniques, and References”, Wiley: New York, 1972; ii) Vogel's Textbook of Practical Organic Chemistry, Pearson Prentice Hall: London, 1989; iii) P. Heinrich Stahl and Camille G. Wernuth 'Handbook of Pharmaceutical Salts', Wiley VCH.

[0127] HPLC analysis.

[0128] Method A: Instrument: Dionex Ultimate 3000 Column: Supelco Ascentis Express C1815 x 4.6 mm 2.7 µm Column temperature: 50℃ Autosampler temperature: Ambient UV wavelength: 275nm Injection volume: 20 μL Flow rate: 1 mL / min Mobile phase A: Water containing 0.1% TFA Mobile phase B: 0.1% TFA in acetonitrile Gradient procedure:

[0129] Method B: In the variant, a Zorbax Extend C18 150x4.6mm 3.5μm column was used for HPLC analysis with the same parameters as described above.

[0130] NMR analysis NMR experiments were performed on a Bruker AVIIIHD spectrometer equipped with a DCH cryopreservation probe, with mass and carbon spectra measured at 500.12 MHz. Experiments were conducted in deuterated DMSO, with each sample prepared to approximately 10 mM concentration. Further analysis was optionally performed in D₂O and / or methanol-d₄. Chemical shifts are expressed in parts per million (ppm, δ units). Coupling constants are expressed in Hz. The abbreviations for multiplicity observed in the NMR spectra are as follows: s (singleton), d (doublet), t (triplet), q (quartet), m (multiplex), br (broad peak).

[0131] UV analysis (for capsule blend homogeneity only)

[0132] Samples were measured using quartz cuvettes with a 10 mm path length. Initial UV scans were performed on a 20 µg / mL salt solution prepared in water, within the 190 nm–400 nm range. Baseline correction was performed using the same solvent. A fixed 280 nm wavelength was primarily used to verify the blending homogeneity of the salt formulation.

[0133] X-ray powder diffraction (XRPD) XRPD analysis was performed on a PANalytical X'pert pro with a PIXcel detector (128 channels), scanning the sample between 3–35° 2θ. The material was gently ground to release any agglomerates and loaded onto a porous plate with a Mylar polymer film for support. The plate was then placed in a diffractometer and analyzed using Cu Kα radiation (α1λ = 1.54060 Å; α2 = 1.54443 Å; β = 1.39225 Å; α1:α2 ratio = 0.5), run using a 40 kV / 40 mA generator in transmission mode (step size 0.0130° 2θ, step time 18.87 s). Data were visualized and images generated using the Highscore Plus 4.7 desktop application (PANalytical, 2017).

[0134] Thermogravimetric / Differential Thermal Analysis (TG / DTA) Approximately 5 mg of material was weighed into an open aluminum pan and placed in a simultaneous thermogravimetric / differential thermal analysis (TG / DTA) instrument, which was then maintained at room temperature. The sample was then heated from 20 °C to 300 °C at a rate of 10 °C / min, and the sample weight was recorded as a function of any differential thermal events (DTA). Nitrogen gas was used as the purge gas at a flow rate of 300 cm⁻¹. 3 / min.

[0135] Differential scanning calorimetry (DSC) Approximately 5 mg of material was weighed into a DSC aluminum pan and non-airtightly sealed with a perforated aluminum cap. The sample pan was then placed in a Seiko DSC6200 (equipped with a cooler) cooled and maintained at 20°C. After obtaining a stable heat flow response, the sample and reference were heated to 180°C at a rate of 10°C / min, and the resulting heat flow response was monitored. Nitrogen gas was used as the purge gas at a flow rate of 50 cm⁻¹. 3 / min.

[0136] Infrared spectroscopy (IR) Infrared spectroscopy was performed on a Bruker ALPHA P spectrometer. Sufficient material was placed at the center of the spectrometer plate, and the spectra were obtained using the following parameters: Resolution: 4cm -1 Background scan time: 16 scans Sample scanning time: 16 scans Data collection: 4000-400cm -1 Result spectrum: transmittance Software: OPUS version 6 Gravimetric vapor adsorption (GVS) Approximately 10–20 mg of sample was placed in a mesh vapor adsorption balance pan and loaded into a Hiden Analytical IGASorp moisture adsorption analyzer. The sample was controlled at 40–90% relative humidity (RH) in 10% increments, held at 25°C at each step until a steady weight was reached (98% of the steps completed, minimum step size 30 min, maximum step size 60 min). After completing the adsorption cycle, the sample was dried to 0% RH using the same procedure, eventually returning to the starting point of 40% RH. Two cycles were performed. The weight change during the adsorption / desorption cycles was plotted to determine the hygroscopic properties of the sample.

[0137] Dynamic vapor adsorption (DVS) Place approximately 10-20 mg of sample in the mesh vapor adsorption balance pan and load the sample into the Intrinsic dynamic vapor adsorption balance of Surface Measurement Systems.

[0138] Samples were subjected to an adsorption / desorption cycle from 40-90% relative humidity (RH) in 10% increments, maintained at 25°C at each step until a steady weight was reached (dm / dt 0.004%, minimum step 30 min, maximum step 500 min). After completing the adsorption cycle, the samples were dried to 0% RH using the same procedure, followed by a second adsorption cycle back to 40% RH. Two cycles were performed. The weight change during the adsorption / desorption cycles was plotted to determine the hygroscopic properties of the samples. Any remaining solids were then analyzed by XRPD.

[0139] Variable humidity X-ray powder diffraction (VH-XRPD) VH-XRPD analysis was performed on a Philips X'Pert pro multi-purpose diffractometer equipped with a humidity chamber. Cu K radiation (α1λ=1.54060Å; α2=1.54443Å; β=1.39225Å; α1:α2 ratio=0.5) was used to scan the sample from 4 to 35.99° 2θ, operating in Bragg-Brentano geometry (step size 0.008° 2θ) with a 40 kV / 40 mA generator. Measurements were taken at 50%, 60%, 70%, 80%, and 90% humidity. The sample was held at 90% humidity for 1 hour and 30 minutes. Measurements were taken at 30-minute intervals.

[0140] Unless otherwise specified, solvents and reagents are purchased and used in the form available from commercial suppliers.

[0141] Example 1 Synthesis and characterization of crystal form 1: N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystalline form of imidazole-2-amine sesquisuccinate Weigh out 514.6 mg in a 20 mL scintillation bottle. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine. 10 mL of methyl ethyl ketone (99% pure) was added to a vial, and the vial was stirred at 40 °C using a temperature control block, stir plate, and magnetic needle. No clear solution was observed, so 5 mL of MEK was added to aid dissolution at 40 °C. A clear solution was obtained; the clear solution at 40 °C was then finely filtered using a PTFE filter (0.45 μm) and a syringe. The filtered solution was added back to a clean 20 mL scintillation vial and reheated to 40 °C. 142.5 mg of succinic acid (1 molar equivalent) was added to the vial. A clear solution was initially observed. After stirring at 40 °C for 30 min, crystallization of the material was observed. The experiment was temperature-cycled for 20 h at 40 °C-5 °C in 3 h cycles. At 5 °C, the slurry was filtered under vacuum using a Buchner flask and funnel with Grade 1 Whatman filter paper (pore size = 11 μm). The wet filter cake was washed with 2.5 mL of cooled (5 °C) MEK solution and dried at 40 °C under reduced pressure for 2 hours. The dried and separated solids were analyzed by XRPD, TG / DTA, and HPLC. The concentration of the filtrate was analyzed by HPLC.

[0142] The separation yield was 78.5%, and the HPLC yield based on the mother liquor concentration was 93.5% (see Table 1).

[0143] Table 1 Preparation results of crystal form 1 succinate

[0144] HPLC (Method A) showed a purity of 99.2%; Rt = 10.65 min ( Figure 7 ).

[0145] The XRPD 2θ diffraction pattern shows that the material is a crystal. Figure 3 When irradiated with a CuKα light source, it contains peaks at diffraction angles of 2θ = 3.8°±0.2° (100%), 10.3°±0.2° (11.3%), 12.4°±0.2° (6.8%), 16.2°±0.2° (23.5%), 17.9°±0.2° (11.9%), 19.8°±0.2° (10.2%), 20.4°±0.2° (13.0%), 23.8°±0.2° (23.3%), and 26.7°±0.2° (9.8%).

[0146] The TG curve shows a 0.4% weight loss from the start of heating up to approximately 160°C due to surface moisture. The DT curve shows an endothermic event starting at approximately 130°C (peaking at 132°C) due to the melting event. Extensive endothermic events were also observed after melting, starting at approximately 209°C (peaking at approximately 228°C); this is related to decomposition. Figure 8 Prepare another batch.N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate was further characterized by HPLC, XRPD, DSC, GVS, FT-IR, NMR and VH-XRPD.

[0147] HPLC (Method A) showed a purity of 98.4%.

[0148] The XRPD 2θ diffraction pattern is the same as that of the aforementioned batch (see...). Figure 12 Top image, for reference Figure 3 ).

[0149] The DSC in the first heating cycle showed a sharp endothermic event starting at approximately 128°C, peaking at 131°C. This was followed by a small exothermic event peaking at 142°C, and then another smaller exothermic event peaking at 147°C. Figure 9 DSC analysis during the first cooling and second heating cycles showed no significant events in the thermograms.

[0150] GVS analysis revealed that the material initially absorbed approximately +0.1 wt% at 70% RH, followed by a sharp increase of approximately +18.9 wt% (approximately 7.1 equivalent water) to 90% RH. During this stage, channel hydrates may have formed. The RH then easily decreased to 70% and gradually decreased to 0% RH. Figure 10 The GVS dynamics diagram can be seen in... Figure 11 It showed a rapid increase in weight between 70-90% RH. XRPD analysis after GVS revealed that the material retained the same crystalline form after exposure to GVS humidity conditions (top: XRPD of the material; bottom: XRPD of the material after GVS).

[0151] FT-IR spectra are shown in Figure 13 middle.

[0152] In DMSO-d6 1 H and quantification 13 The C NMR spectra are shown below. Figure 14 and Figure 15 middle.

[0153] 1H NMR spectrum (500.12 MHz-DMSO-d6): δ (ppm): 7.13 (2H dd J1=6Hz, J2=3Hz), 6.87 (2Hdd J1=6Hz, J2=3Hz), 6.71 (1H br s), 3.30 (2H t J=7Hz), 2.42 (6H m, 2.33 4H dt J1=16Hz, J2=7Hz), 2.04 (4H d J=7Hz), 1.74 (2H quintet J=7Hz), 1.67 (2H septet J=7Hz), 1.52 (2H quintet J=7Hz), 0.85 (12H d J=7Hz) ppm.

[0154] Quantitative 13 C10 NMR spectra (121.16 MHz-DMSO-d6): δ (ppm): 174.3, 156.0, 119.5, 111.9, 63.956.1, 55.6, 53.0 (d), 29.8, 26.8, 26.6, 24.2, 21.3. Quantitative analysis. 13 C10 NMR confirmed the stoichiometry and successful formation of sesquisuccinate.

[0155] VH-XRPD up to 90% RH found that sesquisuccinate crystal form 1 maintained the same crystalline form after exposure to 90% RH for up to 1 hour and 30 minutes, with no change in crystallinity or form.

[0156] 7-day stability test The 7-day stability test was performed as follows: 15 mg of salt was weighed into a 32 mL vial. The vial was then placed at 40°C / 75% RH, 80°C, and under ambient light. After one week, the solid was analyzed for purity by XRPD and HPLC (Method A).

[0157] XRPD analysis of the recovered solids after a 7-day stability test (at 40°C / 75% RH, 80°C and ambient light) showed no change in crystallinity or form after exposure to these conditions. Figure 16 ).

[0158] HPLC analysis (Method A) of the solids recovered from the 7-day stability test showed no change in purity under each stability condition. These values ​​are listed in Table 2.

[0159] Table 2 HPLC purity value of sesquisuccinate crystal form 1 - 7-day stability test

[0160] Solubility test Solubility assessment in unbuffered water showed that the material was highly soluble, with values ​​>500 mg / mL. pH measurements yielded a value of 5.28.

[0161] Weigh out approximately 20mg N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate crystals were placed in 1 to 3 2 mL vials. 100 μL of appropriate buffer solution was added to form a slurry. The results are listed in Table 3.

[0162] Table 3 : Solubility value of sesquisuccinate crystal form 1

[0163] Synthesis scale-up Two additional batches of this sesquisuccinate crystal form 1 were synthesized using the same experimental conditions, starting with 20 g and 79 g of free base. The purity of these batches, as determined by HPLC (Method A), was ≥ 99%, with yields of 69% and 73%, respectively. Overlapping XRPD 2θ diffraction patterns for all batches (514 mg, 20 g, and 79 g scales) are shown below. Figure 17 As shown in the image.

[0164] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine, sesquisuccinate crystal form 1 can be obtained repeatedly in multi-gram scales using a one-pot salt formation / crystallization process with good yields. Sesquisuccinate crystal form 1 is very stable (in both purity and polymorphic form), highly crystalline, highly soluble, non-hygroscopic at up to 90% relative humidity, and stable under stress conditions. Therefore, this product is a suitable API and is particularly beneficial for solid dosage forms.

[0165] Example 2 Synthesis and characterization of crystal form 2: N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Crystalline form of imidazole-2-amine trisuccinate 10mg N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazol-2-amine was weighed into a 1.5 mL vial and a minimum amount of ethanol was added to ensure complete dissolution of the free base. Two equivalents of succinic acid were added using a 0.5 M stock solution (596.41 mg succinic acid dissolved in 10 mL ethanol). The vial was cyclically heated between ambient temperature and 40 °C (4 hours per cycle) with stirring for 6 days. A precipitate was observed; the solid was separated by centrifugation and filtration, and then vacuum dried at 40 °C for approximately 24 hours. The solid was analyzed by HPLC, XRPD, TG / DT, DSC, DVS, FT-IR, NMR, and VH-XRPD.

[0166] HPLC (Method A) showed a purity of 99.9%.

[0167] The XRPD 2θ diffraction pattern shows that the material is crystalline (see [link]). Figure 4 When irradiated with a CuKα light source, it contains peaks at diffraction angles of 2θ = 5.3°±0.2° (100%), 15.0°±0.2° (26.7%), 15.3°±0.2° (57.4%), 16.8°±0.2° (25.3%), 17.9°±0.2° (92.3%), 20.5°±0.2° (30.1%), 21.2°±0.2° (25.5%), 23.9°±0.2° (24.2%), 24.3°±0.2° (65.3%), and 26.6°±0.2° (31.5%).

[0168] TG curves from TG / DT analysis (see) Figure 18 The results show no mass loss up to 150°C, indicating that the material contains no residual solvent. Mass loss associated with salt dissociation was observed at temperatures above 150°C. The DT curve shows the endothermic events associated with coupling dissociation / melting in the salt form, starting at approximately 155.1°C (peak at 159.1°C).

[0169] DSC thermal analysis chart (see...) Figure 19 The results show that trisuccinate crystal form 2 begins to melt at approximately 155.8 °C (peak at 157.7 °C).

[0170] Performed in MeOH-d4 1 ¹H NMR confirmed that the salt contains 3 equivalents of succinic acid (see [reference]). Figure 20 ).

[0171] 1H NMR spectrum (500.12 MHz-MeOH-d4): δ (ppm): 7.28 (2H dd J1=6Hz, J2=3Hz), 7.10 (2Hdd J1=6Hz, J2=3Hz), 3.48 (2H t J=7Hz), 2.78 (6H br s), 2.67 (7H m), 2.56 (12H s), 2.41 (4H d J=7Hz), 1.95 (2H quintet J=7Hz), 1.87 (2H septet J=7Hz), 1.79 (2H sextet J=7.4Hz), 1.33 (5H br m), 0.97 (12H d J=6.7Hz) ppm.

[0172] DVS analysis revealed that the material absorbed approximately +0.1 wt% at 70% RH, followed by a sharp increase of approximately +9.92 wt% (approximately 4.3 equivalents of water) up to 90% RH. Then, it easily decreased to 70% RH and gradually dropped to 0% RH. Figure 21 The DVS isotherm plot is related to the Type I isotherm, indicating that the structure does not change throughout the DVS cycle, as shown in the DVS kinetic plot. Figure 22 As confirmed by [reference needed], no significant changes in quality were observed. Post-DVS XRPD analysis showed that the material retained the trisuccinate crystal form.

[0173] 7-day stability studies showed that crystal form 2 was stable at 50°C, 80°C, and under ambient light conditions. Solubility in different buffer solutions is reported in Table 4.

[0174] Table 4 Solubility value of trisuccinate crystal form 2

[0175] N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine trisuccinate crystal form 2 was obtained by one-pot salting / crystallization with good yield. Trisuccinate crystal form 2 is very stable (in both purity and polymorphic form), highly crystalline, highly soluble, non-hygroscopic at up to 90% relative humidity, and stable under stress conditions. Therefore, this product is a suitable API and is particularly beneficial for solid dosage forms.

[0176] Example 3 :contain N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dPreparation and stability of capsules containing imidazole-2-amine sesquisuccinate crystal form 1 Preparation of formulation blends Add 7.5g of Avicel ® PH101 was weighed into a 220mL cylindrical drum, and then 15g was weighed. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate crystal form 1, finally weighed 7.5g of Avicel ® PH101. Crystal form 1 and Avicel are added in this manner. ® PH101 excipient improved the mixing of the two compounds. The final blend was a 50:50 w / w crystal form 1:Avicel. ® PH101. The formulation was mixed using a TorPac powder mixer / blender by rotating the drum at a fixed speed of 55-65 rpm for 5 minutes, then counter-rotating every 60 seconds. After 5 minutes, a subsample was removed, and an aqueous solution equivalent to 20 μg / mL of crystal form 1 was prepared. The content of crystal form 1 in the blend was determined by UV analysis relative to a standard of the same salt and concentration. Mixing continued until a homogeneous blend was obtained. Since the excipient is insoluble in water, the sample solution was filtered through a 0.45 μm PTFE syringe filter prior to analysis. This standard was used to confirm that the selected membrane did not adversely affect the recovery of the analyte. After obtaining a homogeneous blend, 100 hard gelatin-sized white / white capsules were filled. The homogeneity of the filled capsules was assessed by weighing 10 filled capsules from each batch. Variations in empty shells were considered insignificant.

[0177] Stability Sample Preparation of Formulation Blends The filled capsules were dispensed into HDPE screw-cap vials, 10 capsules per vial, and stored. Four vials were stored under each condition: RT and 40°C / 75% RH. Ten capsules were used for the initial time point and for determining weight homogeneity. At each time point, five capsules were cut open and the contents were dispensed into 20 mL scintillation vials. The powder was then mixed with a spatula and sampled as needed for HPLC (Method B) and XRPD analysis.

[0178] Uniformity of blends When the concentrations of the three simultaneous subsamples are all within 5%, blend homogeneity is established. Sesquisuccinate crystal form 1 requires only 30 minutes of mixing to obtain a uniform powder.

[0179] Table 5 : Homogeneity value of sesquisuccinate crystal form 1 blend

[0180] In contrast, it was found that the disulfate form B blend required up to 60 minutes to form small aggregates during mixing. Applying excessive shear force with disulfate form B is necessary to disrupt the aggregates and accelerate (i.e., reduce to 30 minutes) the mixing process, but may also adversely affect crystallinity.

[0181] Capsule homogeneity: weight of filled capsules Due to the free-flowing nature of the blend, powder formulations are easily filled into capsules, and suitable capsule uniformity is achieved.

[0182] Table 6 The weight of capsules filled with the sesquisuccinate crystal form 1 blend

[0183] HPLC stability determination No change in purity was found throughout the stable storage period. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate crystal form 1 is retained during the entire 4-week storage of the capsule at room temperature or 40°C / 75% RH.

[0184] Table 7 HPLC purity of capsules filled with sesquisuccinate crystal form 1 blend

[0185] XRPD stability analysis No change in crystal form was observed throughout the stable storage, N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine sesquisuccinate crystal form 1 retains its crystal form during the entire 4-week storage period of the capsules at room temperature or 40°C / 75%RH (see [link]). Figure 23 and 24 ).

[0186] Will N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ dImidazole-2-amine sesquisuccinate crystal form 1 is effectively blended with microcrystalline cellulose excipients to provide a free-flowing powder that is easily dispersed in capsules. As the purity and crystalline form of the API remain unchanged, stability is achieved for 4 weeks at RT and 40°C / 75% RH.

[0187] Biological Examples Aβ 1-42 Effects on cortical neuron damage Cortical neuron culture Modified as described by Callizot et al. in J Neurosci Res. 2013, 91: 706-716 to include microglia, rat cortical neurons cultured with microglia were cultured. Pregnant female rats (Rats Wistar; Janvier Labs France) at 15 days of gestation were euthanized by cervical dislocation after deep CO2 anesthesia. The fetuses were then collected and immediately placed in ice-cold L15 Leibovitz medium containing 2% penicillin (10,000 U / ml) and streptomycin (10 mg / ml) solution (PS) and 1% bovine serum albumin (BSA). The cortex was treated with trypsin-EDTA solution at 37°C for 20 minutes at a final concentration of 0.05% trypsin and 0.02% EDTA. Dissociation was terminated by adding Dulbecco modified Eagle medium (DMEM) containing 4.5 g / L glucose, grade II DNAse I (final concentration 0.5 mg / ml), and 10% fetal bovine serum (FCS). Cells were mechanically dissociated three times through the channel of a 10 mL pipette tip. The cells were then centrifuged at 515 x g for 10 min at 4 °C. The supernatant was discarded, and the pellet was resuspended in a defined composition medium consisting of Neurobasal medium containing 2% B27 supplementation solution, 2 mmol / L L-glutamine, 2% PS solution, 10 ng / mL brain-derived neurotrophic factor (BDNF), 2% heat-inactivated horse serum, 2% heat-inactivated FCS, 1 g / L glucose, 1 mM sodium pyruvate, and 100 μM non-essential amino acids. Viable cells were counted using a Neubauer cell counter using a trypan blue exclusion assay. Cells were seeded at a density of 45,000 cells per well in 96-well plates pre-coated with poly-L-lysine (for immunostaining) and cultured at 37 °C in an air (95%)-CO2 (5%) incubator. The medium was changed every 2 days.

[0188] Test compounds and human Aβ 1-42 Exposure After 11 days of culture, cortical neurons were damaged with Aβ solution (see below). Aβ1-42 The preparation was performed according to the procedure described by Callizot et al., 2013. In short, Aβ... 1-42 The peptide was dissolved at an initial concentration of 40 μM in the culture medium determined above. The solution was gently stirred in the dark at 37°C for 3 days and then appropriately diluted in control culture medium to the desired concentration (5 μM Aβ containing 0.5 μM Aβ oligomer (AβO) as measured by Western blotting). 1-42 Use immediately after formulation. The control medium consists of the defined medium composition (as described above). Dissolve the test compound in water and mix it in the medium. Mix the compound with Aβ. 1-42 The formulation was administered in combination with primary cortical neurons cultured with microglia for 72 hours.

[0189] Immunostaining After permeabilization with 0.1% saponin, cells were blocked with PBS containing 1% fetal bovine serum for 15 minutes. Then, the cells were incubated with the following substances for 2 hours: a) For MAP-2 neurons: Chicken polyclonal antibody (Ab) against microtubule-associated protein 2 (MAP-2), diluted 1 / 1000 in PBS containing 1% fetal bovine serum and 0.1% saponin. This antibody specifically binds to neurons and synapses to investigate neuronal cell survival and synaptic networks.

[0190] b) For AT100 Tau phosphorylation: mouse anti-tau monoclonal antibody, phosphorylated at Thr212 and Ser214 sites (AT100), diluted 1 / 400 in PBS containing 1% fetal bovine serum and 0.1% saponin.

[0191] c) For OX-41 microglia: Rabbit anti-SIRPα / CD172a (OX-41) polyclonal Ab, diluted 1 / 400 in PBS, containing 1% fetal bovine serum and 0.1% saponin. This Ab specifically binds to microglia, allowing for the assessment of their activation.

[0192] Alexa Fluor 350 donkey anti-chicken IgG, Alexa Fluor 488 goat anti-mouse IgG, and Alexa Fluor 568 goat anti-rabbit IgG were diluted 1 / 400 in PBS containing 1% FCS and 0.1% saponin and exposed to room temperature for 1 hour.

[0193] For each condition, ImageXpress (Molecular Devices) automatically captured 30 images per well at 20x magnification. All images were generated using the same acquisition parameters. Analysis was performed automatically using the Custom Module Editor (Molecular Devices).

[0194] The following readings were studied: - Neuron survival: Total number of neurons (MAP-2 staining) - Neural synaptic network (MAP-2 staining, in μm) - Co-localization area (overlap μm) of phosphorylated Tau:AT100 and MAP-2 within neurons 2 ) - Microglia activation: Total microglia area (OX41 staining, in μm) 2 count) Quantitative analysis of extracellular granule protein precursors Following the manufacturer's recommendations, the level of PGRN in the culture medium was determined by ELISA (Particle Progenitor (Rat) ELISA lit, Adipogen, Coger). In short, 100 μL of supernatant was added directly to the well strip at 37°C for 1 hour. Then, 100 μL of detection antibody was added at 37°C for 1 hour, followed by 100 μL of HRP-labeled streptavidin at 37°C for 1 hour. Finally, 100 μL of TMB substrate solution was added at room temperature for 10 minutes. Optical density (OD) was assessed spectrophotometrically at 450 nm using a Glomax instrument (Promega). Peroxidase activity was proportional to the concentration of PGRN.

[0195] Statistical analysis Results are expressed as a percentage of the control. All values ​​are shown as the mean + / - SEM (standard error of the mean) for 4–6 wells under each condition. Graphical and statistical analyses (ANOVA, followed by Dunnett or Fisher t-test) were performed under different conditions using Graphpad Prism software version 7.04. p < 0.05 is considered significant.

[0196] result Neuron survival Compared with the control group, Aβ 1-42 Damage significantly reduces the survival of cortical neurons. Figure 25 A). Sesquisuccinate crystal form 1 is neuroprotective at the highest doses (10 nM-100 nM).

[0197] Neural synaptic network :Aβ 1-42 The total length of the neural synaptic network is significantly reduced after injury. Figure 25 B). In the presence of the highest dose of sesquisuccinate crystal form 1 (10 nM-100 nM), the neural synaptic network is longer.

[0198] AT100 In applying Aβ 1-42 At that time, a significant increase in AT100 was observed in MAP-2 cortical neurons. Figure 25 C). Sesquisuccinate form 1 significantly reduces Tau hyperphosphorylation at doses above 1 nM.

[0199] Microglia activation As expected, Aβ levels were lower than those in the control group. 1-42 Damage triggers microglial cell activation ( Figure 26 A). At the studied dose, sesquisuccinate crystal form 1 reduced microglial cell activation.

[0200] extracellular PGRN : In Aβ 1-42 In the presence of PGRN, the level in the supernatant was significantly lower than that in the control group. Sesquisuccinate crystal form 1 significantly increased the level of extracellular PGRN at a concentration of 10 nM. Figure 26 B).

[0201] Consistent with these results, it can be concluded that sesquisuccinate crystal form 1 is capable of a) being neuroprotective, b) protecting neural synaptic networks, c) reducing Tau hyperphosphorylation, d) alleviating microglial activation and thus reducing neuroinflammation, and e) promoting the release of granule protein precursor neurotrophic factor from cultured cell supernatants. Therefore, the succinate of the present invention can be used to treat neurodegenerative diseases, particularly tau protein diseases and Parkinson's disease.

[0202] Effects of MPP + or rotenone on midbrain neurons Midbrain neuron culture Rat dopaminergic neurons were cultured as described by Visanji et al. in FASEB J. 2008, 22(7): 2488-2497 and Callizot et al., in PlosOne 2019: https: / / doi.org / 10.1371 / journal.pone.0215277. In short, pregnant female rats (Wistar) at 15 days of gestation were euthanized by deep anesthesia and cervical dislocation in a CO2 chamber. The midbrain obtained from 15-day-old rat embryos (Janvier, France) was dissected under a microscope. The embryonic midbrain was removed and placed in Leibovitz (L15) ice-cold medium containing 2% penicillin-streptomycin (PS) and 1% BSA. The ventral portion of the midbrain, i.e., the region of the developing midbrain rich in dopaminergic neurons, was used for cell preparation. Midbrain cells were dissociated by trypsin digestion at 37°C for 20 minutes (final concentration: 0.05% trypsin and 0.02% EDTA). The reaction was terminated by adding DMEM containing grade II DNAse I (0.5 mg / mL) and 10% FCS. Cells were then mechanically separated three times using 10 mL pipettes. Cells were then centrifuged at 180 x g for 10 min on a layer of BSA (3.5%) in L15 medium at +4°C. The supernatant was discarded, and the cell pellet was resuspended in a defined-component medium supplemented with B27 (2%), L-glutamine (2 mM), and 2% PS solution, as well as 10 ng / mL BDNF and 1 ng / mL glial-derived neurotrophic factor (GDNF). Viable cells were counted using a trypan blue exclusion assay in a Neubauer cell counter. Cells were seeded at a density of 40,000 cells / well in 96-well plates (pre-coated with poly-L-lysine) and incubated in a humidified incubator at 37°C in a 5% CO2 / 95% air atmosphere. Half of the culture medium was replaced with fresh medium every 2 days.

[0203] For 96-well plates, use only 60 wells. Do not use the first and last rows and columns of wells (to avoid any edge effects) and fill them with sterile water.

[0204] Test compounds and mitochondrial stress Medium: Culture medium (0.1% PBS).

[0205] Pretreatment: On day 4, sesquisuccinate crystal form 1 was dissolved in the culture medium and added to the culture for 48 hours.

[0206] MPP +Damage: On day 6 and 48 after incubation of sesquisuccinate crystal form 1, or immediately after incubation, MPP+ was added to a final concentration of 4 μM and diluted for 48 hours in control medium still containing the compound.

[0207] Rotenone damage: Rotenone was added to a final concentration of 4 μM on day 6 and day 48 after incubation of sesquisuccinate crystal form 1, or immediately after incubation, and diluted for 24 hours in control medium in which the compound was still present.

[0208] Immunostaining Following injury (24 or 48 hours), the cell culture supernatant was removed, and cells were fixed for 20 minutes at room temperature in PBS solution containing 4% paraformaldehyde (pH 7.3). Cells were washed twice in PBS. Cell membrane permeabilization and non-specific binding site blocking were performed for 15 minutes at room temperature using PBS solution containing 0.1% saponin and 1% FCS. Cells were then incubated with the following: a) For tyrosine hydroxylase (TH): Monoclonal anti-TH antibodies generated in mice were diluted 1 / 10000 in PBS containing 1% FCS and 0.1% saponin and incubated at room temperature for 2 hours.

[0209] b) For α-synuclein: The polyclonal anti-α-syn antibody produced in rabbits was diluted 1 / 200 in PBS containing 1% FCS and 0.1% saponin and incubated at room temperature for 2 hours.

[0210] Alexa Fluor 488 goat anti-mouse IgG and Alexa Fluor 568 goat anti-rabbit IgG, both diluted 1 / 400 in PBS containing 1% FCS and 0.1% saponin, were exposed to the primary antibody for 1 hour at room temperature. Cell nuclei were labeled with Hoechst dye (1 / 1000).

[0211] For each condition, 20 images (representing the entire well area) were automatically captured using ImagExpress® (Molecular Devices) at 10x magnification with the same acquisition parameters. The images were then automatically analyzed using MetaXpress® (Molecular Devices).

[0212] The following readings were studied: - Neuron survival: Total number of TH neurons (TH staining) - Neural synaptic network: Total length of neural synaptic network of TH-positive neurons (μm) - α-syn accumulation (overlap area between TH and α-syn staining, μm) 2 ) Statistical analysis Results are expressed as a percentage of the control. All values ​​are shown as the mean + / - SEM (standard error of the mean) for 4–6 wells under each condition. Statistical analysis of the different conditions (ANOVA, followed by Fisher's LSD test) and graphs was performed using Graphpad Prism software version 7.04. p < 0.05 was considered significant.

[0213] result Sesquisuccinate crystal form 1 pair MPP + The effect of damaged TH(+) neurons Neuron survival Compared with the control group, MPP + Damage significantly reduces the survival of dopaminergic (TH-positive) neurons. Sesquisuccinate crystal form 1 has neuroprotective effects at a dose of 50 nM. Figure 27 A).

[0214] Neural synaptic network MPP + The total length of the neural synaptic network was significantly reduced after injury. A 50 nM dose of sesquisuccinate crystal form 1 significantly improved the integrity of the neural synaptic network. Figure 27 B).

[0215] α-syn accumulation In applying MPP + A significant increase in α-syn accumulation was observed in dopaminergic neurons. This accumulation was significantly reduced after pre-incubation with sesquisuccinate form 1 at all tested concentrations. Co-incubation with 50 or 100 nM sesquisuccinate form 1 significantly reduced α-syn accumulation. Figure 27 C).

[0216] Effects of sesquisuccinate crystal form 1 on rotenone-damaged TH(+) neurons Neuron survival Compared to the control group, rotenone induced significant loss of dopaminergic neurons. When pre-incubated, sesquisuccinate form 1 exhibited significant neuroprotective effects at doses of 10 and 50 nM. However, when administered directly with rotenone, it significantly increased neuronal survival at all concentrations. Figure 28 A).

[0217] Neural synaptic network Rotenone damage significantly reduced the integrity of the neural synaptic network. Pretreatment and co-treatment with sesquisuccinate crystal form 1 significantly improved the neural synaptic network at all doses ranging from 10 to 100 nM. Figure 28 B).

[0218] α-syn accumulationRotenone significantly increases the accumulation of α-synuclein in dopaminergic neurons. Pathological accumulation was significantly reduced when sesquisuccinate form 1 was pre-incubated with or directly administered with rotenone (10-100 nM). Notably, co-administration of 100 nM sesquisuccinate form 1 prevented the accumulation of α-synuclein. Figure 28 C).

Claims

1. N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine succinate and its pharmaceutically acceptable solvates.

2. The succinate of claim 1, having formula II Formula II Where x is 1-4, and its pharmaceutically acceptable solvates.

3. The succinate and its pharmaceutically acceptable solvates according to claim 2, characterized in that, x is 1.4-3.

1.

4. The succinate and its pharmaceutically acceptable solvates according to claim 2, characterized in that, x is 1.4-1.

6.

5. The succinate and its pharmaceutically acceptable solvates according to claim 2, characterized in that, x is 1.

5.

6. The succinate and its pharmaceutically acceptable solvates according to claim 2, characterized in that, x is 2.9-3.

1.

7. A pharmaceutical composition comprising a succinate or a pharmaceutically acceptable solvate thereof according to any one of claims 1-6, and at least one pharmaceutically acceptable carrier, diluent, excipient and / or adjuvant.

8. Used for preparation N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d A method for determining the crystal form of imidazole-2-amine sesquisuccinate, the method comprising the following steps: - Step 1: Put N -(3-(4-(3-(diisobutylamino)propyl)piperazin-1-yl)propyl)-1 H -benzo[ d Imidazole-2-amine dissolves in an organic solvent; - Step 2: Heat the reaction medium to temperature B, where temperature B is defined as 30℃-50℃; - Step 3: Add approximately an equimolar amount of succinic acid; - Step 4: Stir the reaction medium at temperature B for 0.25-4 hours; - Step 5: Cycle the reaction medium between temperature A and temperature B for 2-4 hours for 10-30 hours, where temperature A is defined as 0-10℃. - Step 6: Cool the reaction medium to temperature A; - Step 7: Filter the reaction medium at temperature A; - Step 8: Wash the filter cake with the organic solvent at temperature A; - Step 9: Dry the filter cake at a temperature of 30-50℃.

9. The succinate or a pharmaceutically acceptable solvate thereof according to any one of claims 1-6, used as a medicine.

10. The succinate or a pharmaceutically acceptable solvate thereof according to any one of claims 1-6, for the treatment and / or prevention of diseases selected from neurodegenerative diseases and diseases in which impaired phosphorylation of Tau protein is observed.

11. The succinate or a pharmaceutically acceptable solvate thereof according to any one of claims 1-6, for use in delaying the onset of diseases selected from neurodegenerative diseases and diseases in which impaired phosphorylation of Tau protein is observed.

12. The succinate or a pharmaceutically acceptable solvate thereof for use according to claim 10 or 11, wherein the disease is tau proteinosis.

13. The succinate or a pharmaceutically acceptable solvate thereof for use according to claim 12, wherein the tau protein disease is selected from Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson's-Guam dementia syndrome, aerobatic granuloma, chronic traumatic encephalopathy, corticobasal degeneration, muscular dementia, diffuse neurofibrillary tangles with calcification, familial British dementia, familial Danish dementia, frontotemporal dementia (FTD), frontotemporal dementia with chromosomal 17-linked Parkinson's disease (FTDP-17), frontotemporal degeneration (FTLD), frontotemporal dementia-granuloma subtype (FTD-GRN), Gerstmann-Sträussler-Scheinker disease, Guadelphia... Oupean Parkinson's disease, Hallewarden-Schpattz disease, inclusion body myositis, multiple system atrophy, Steinert's myotonic dystrophy, type II myotonic dystrophy, neurodegeneration with iron buildup in the brain, Niemann-Pick disease type C, Non-Guamanian motor neuron disease with neurofibrillary tangles, Paget's disease, Pick's disease, post-encephalitis Parkinson's syndrome, progressive subcortical glial proliferation, progressive supranuclear palsy (PSP), SLC9A6-related intellectual disability, subacute sclerosing panencephalitis, tangles dementia, multi-infarct dementia, ischemic stroke, chronic traumatic encephalopathy (CTE), traumatic brain injury (TBI), stroke, and white matter tau proteinosis with globular glial inclusions.

14. The succinate or a pharmaceutically acceptable solvate thereof for use according to claim 10 or 11, wherein the disease is Parkinson's disease.

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