TREATMENT OF CNS DISEASES WITH sGC STIMULATORS

Abstract

This invention relates to the use of stimulants of soluble guanylate cyclase (sGC), pharmaceutically acceptable salts thereof, and pharmaceutical formulations or dosage forms comprising them, alone or in combination with one or more additional agents, for the treatment of various CNS diseases, wherein increased sGC stimulation, or increased concentrations of nitric oxide (NO) or cyclic 3',5'-guanosine monophosphate (cGMP), or both, or upregulation of the NO-sGC-cGMP pathway, is suitable. The compounds suitable for use in the methods of this invention are those of Formula I or their pharmaceutically acceptable salts.

Description

Technical Field

[0001] This invention relates to the use of soluble guanylate cyclase (sGC) stimulants, pharmaceutically acceptable salts thereof, and pharmaceutical formulations or dosage forms containing them, alone or in combination with one or more additional agents, for the treatment of various CNS diseases, wherein increased sGC stimulation, or increased concentrations of nitric oxide (NO) or cyclic 3',5'-monophosphate guanosine (cGMP), or both, or upregulation of the NO-sGC-cGMP pathway is preferred. Prior Technology

[0002] sGC is the primary receptor for NO in vivo. Upon binding to sGC, NO activates its catalytic domain, leading to the conversion of guanosine-5'-triphosphate (GTP) into the second messenger, cGMP. Increased levels of cGMP further regulate the activity of downstream effectors, including protein kinases, phosphodiesterases (PDEs), and ion channels. In vivo, NO is synthesized from arginine and oxygen via various nitric oxide synthases (NOS) and the sequential reduction of inorganic nitrates. Experimental and clinical evidence indicates that decreased NO concentration, reduced NO bioavailability, and / or decreased responsiveness to endogenously produced NO contribute to the development of numerous diseases. sGC stimulators are heme-dependent agonists of the sGC enzyme, synergistically acting with varying amounts of NO to enhance the enzymatic conversion of GTP to cGMP. sGC stimulators are clearly distinguished from another class of known sGC activators—NO-independent, heme-independent agonists—and are structurally unrelated.

[0003] Therapies that improve or restore sGC function offer significant advantages over existing alternative therapies that target the NO-sGC-cGMP pathway or otherwise benefit from its upregulation. There is an urgent need to develop novel and safe therapies for patients with functional impairments or downregulated NO-sGC-cGMP pathways.

[0004] sGC stimulants, capable of crossing the blood-brain barrier (BBB) ​​and penetrating into the central nervous system (CNS), offer additional benefits for the treatment of CNS diseases. The sGC stimulants described in this article are suitable for treating CNS diseases due to their ability to cross the BBB and activate targets in the brain. There is an urgent need to develop additional novel therapeutic agents for the treatment of CNS diseases. The use of sGC stimulation in the CNS represents a novel mechanism of action for treating these diseases that has only recently begun to be investigated, as previously, sGC stimulants with CNS-penetrating properties were lacking. Summary of the Invention

[0005] This invention is based on the discovery that the compounds disclosed herein are sGC stimulants capable of penetrating the blood-brain barrier (BBB) ​​and therefore suitable for treating CNS diseases. Compounds with relevant structural features, specifically 4-OH substituents on a pyrimidine ring, were previously known only as synthetic intermediates for preparing sGC stimulants with 4-amino substituents on a pyrimidine ring. Compounds of this class in previous disclosures did not have medical applications. Unexpectedly, the compounds of this invention have been found to possess strong sGC stimulating activity, penetrating the BBB, increasing cGMP concentration in the brain, and exhibiting CNS activity in both in vitro and in vivo analyses.

[0006] In a first embodiment, the present invention relates to a method for treating CNS diseases, CNS health conditions, or CNS symptoms in an individual in need, comprising administering, alone or in combination, a therapeutically effective amount of a compound represented by formula I: ; Or a medically acceptable salt thereof, wherein: JC series is selected from the following groups: hydrogen, halogen, C1-6 alkyl, and C1-6 fluoroalkyl with 1 to 3 fluorine atoms substituted; X is either N or C (J C1); J C1 series are selected from the following groups: hydrogen, halogens, C1-6 alkyl groups and C1-6 fluoroalkyl groups substituted with 1 to 3 fluorine atoms; Each JB is independently selected from the group consisting of: hydrogen, halogen, C1-6 alkyl, and C1-6 fluoroalkyl with 1 to 3 fluorine atoms substituted; The JD series is selected from the group consisting of: hydrogen, halogens, C1-6 alkyl groups and C1-6 fluoroalkyl groups substituted with 1 to 3 fluorine atoms; and n is an integer selected from 0, 1, 2, 3, or 4. In a second embodiment, the present invention relates to a method for treating a CNS disease, CNS health condition, or CNS symptom in an individual in need, comprising administering, alone or in combination, a therapeutically effective amount of a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof to the individual.

[0007] In a third embodiment, the present invention further relates to the use of a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the manufacture of an agent for treating CNS diseases, CNS health conditions or CNS disorders in an individual in need.

[0008] In a fourth embodiment, the present invention further relates to a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the treatment of CNS diseases, CNS health conditions or CNS disorders in an individual in need. Simple Explanation of the Diagram

[0009] Figure 1 shows the effect of compound I-14 on the change in MAP relative to baseline (ΔBMAP) in male normotensive rats. Figure 2 shows the effect of compound I-20 on ΔBMAP in male normotensive rats. Figure 3 shows the concentrations of compound I-14 in the STR and HIPP regions of the brain of adult male Spurgeon-Dolly rats after administration of PO (3 mg / kg) at T = 0 min. Data are presented as mean ± SEM, N = 5. Figure 4 shows the concentrations of cGMP in rat CSF at 1, 2, and 6 hours after administration of a single oral dose of compound I-20 (1 mg / kg, 3 mg / kg, or 10 mg / kg). Figure 5 shows the concentrations of cGMP in rat CSF at 1, 2, and 6 hours after administration of a single oral dose of compound I-14 (1 mg / kg, 3 mg / kg, 10 mg / kg, or 30 mg / kg). Figure 6 illustrates the cognitive effects of compound I-14 in a macaque model of long-term low-dose MPTP-induced cognitive deficits due to Parkinson's disease. SDR efficacy was significantly reduced after long-term low-dose MPTP exposure (**P<0.01). Following mediator administration, SD (simple discrimination) and SDR (simple discrimination reversal) efficacy were identical to baseline MPTP efficacy. SDR efficacy was significantly improved after compound I-14 administration (**P<0.01) and worsened during clearance (**P<0.01 compared to drug efficacy). Figure 6A shows mean ± SEM efficacy; and Figure 6B is a scatter plot of individual data using mean ± SEM. N = normal, before MPTP; WO = clearance. Implementation

[0010] Related applications

[0011] This application claims priority to U.S. Provisional Application No. 63 / 177,023, filed April 20, 2021, and U.S. Provisional Application No. 63 / 229,251, filed August 4, 2021. The entire contents of each of the foregoing applications are incorporated herein by reference.

[0012] Reference will now be made in detail to certain embodiments of the present invention, examples of which are illustrated in the accompanying structures and formulas. While the invention will be described in conjunction with the enumerated embodiments, it should be understood that it is not intended to limit the invention to those embodiments. Rather, the invention is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of the invention as defined in the claims. The invention is not limited to the methods and materials described herein, but includes any methods and materials similar to or equivalent to those described herein that may be used in the practice of the invention. In the event that one or more of the incorporated references, patents, or similar materials (including, but not limited to, defined terminology, usage of terminology, described techniques, etc.) differ from or conflict with this application, this application shall prevail.

[0013] [, Definitions and General Terms , ] [, , ]

[0014] For the purposes of this invention, chemical elements are identified according to the periodic table, CAS edition, and Handbook of Chemistry and Physics, 75th edition, 1994. Furthermore, the general principles of organic chemistry are described in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," 5th edition, Smith, MB and March, J. (eds.), John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.

[0015] When one or more positions of a structure can be substituted by one or more substituents selected from a specified group or list, the substituents at each position may be "independently selected" to be equal or identical at each position and for each example, unless otherwise specified. For example, if a phenyl group is substituted by two instances of R 100, and each R 100 is independently selected from a halogen and a methyl group, it means that each instance of R 100 is selected from a halogen or a methyl group; for example, one R 100 may be fluorine and one may be methyl, or both may be chlorine, etc. Similarly, if the substituted atom is bonded to at most one hydrogen atom (e.g., CH 3 or NH 2), the substituents may be "independently selected" to be equal or identical at each position and for each example, unless otherwise specified. For example, if a methyl group (e.g., CH 3) is substituted by two instances of R 100 and each R 100 is independently selected from a halogen and a methyl group, then each instance of R 100 is selected from a halogen or a methyl group; for example, one R 100 may be fluorine and the other may be methyl (e.g., CHF(CH 3), or both may be chlorine (e.g., CHCl 2), etc.

[0016] The selection and combination of substituents contemplated in this invention are merely those selections and combinations that form stable or chemically viable compounds. Such selections and combinations will be obvious to those skilled in the art and can be determined without improper experimentation. As used herein, the term "stable" means a compound that remains substantially unchanged when subjected to conditions permissible for its production, detection, and, in some embodiments, recovery and purification, and for use in one or more of the purposes disclosed herein. Chemically viable compounds are those that can be prepared by those skilled in the art based on the disclosures herein (and, where necessary, relevant knowledge of the art).

[0017] Unless otherwise stated, all tautomer forms of the compounds of this invention are also within the scope of this invention.

[0018] In one embodiment, the invention may include replacing hydrogen with deuterium (i.e., 2H), thereby obtaining certain therapeutic advantages due to greater metabolic stability (e.g., prolonged in vivo half-life or reduced dose requirement) and is therefore preferred in some cases. The deuterated compounds of the invention can generally be prepared by a procedure similar to the processes described below and / or the procedures disclosed in the examples, by replacing the deuterated reagent with an undeuterated reagent.

[0019] As used herein, the term "alkyl" as used, such as "alkyl chain" or "alkyl", refers to a saturated unbranched (e.g., straight-chain) or branched monovalent hydrocarbon group. "Cxyalkyl" (where x and y are two distinct integers, both different from 0) is an alkyl chain containing a number of carbon atoms between x and y (including the endpoints). For example, C1-6 alkyl refers to any alkyl group containing a number of carbon atoms between 1 and 6 as defined above. Examples of alkyl groups include (but are not limited to) methyl (i.e., C1 alkyl), ethyl (i.e., C2 alkyl), n-propyl (C3 alkyl), isopropyl (different C3 alkyl), n-butyl, isobutyl, secondary butyl, tertiary butyl, pentyl, hexyl, heptyl, octyl, and the like. In some embodiments, the alkyl group is C1-4 alkyl. In some embodiments, the alkyl group is C1-3 alkyl or C1-2 alkyl. In other embodiments, the alkyl group is methyl or ethyl.

[0020] As used herein, the term "fluoroalkyl" means an alkyl group as defined above, wherein one or more of the hydrogen atoms attached to the chain carbon atoms at any one or more carbon atoms of the alkyl group have been replaced by fluorine. For example, a fluoroalkyl group substituted with 1 to 3 fluorine atoms is an alkyl group in which 1 to 3 hydrogen atoms at any position on the same or different carbon atoms of the alkyl chain have been replaced by fluorine atoms.

[0021] As used herein, the terms "halogen" or "halogen group" mean F, Cl, Br, or I. In some embodiments, the halogen group is F or Cl. In other embodiments, the halogen group is F.

[0022] The term "hydroxyl" refers to -OH.

[0023] The compounds of this invention are defined herein by their chemical structure and / or chemical name. When a compound is mentioned by its chemical structure and chemical name and there is a conflict between the chemical structure and chemical name, the chemical structure determines the identity of the compound.

[0024] [, , ] [, Examples of compounds and compositions , ] [, , ] This invention relates to the medical use of a compound of formula I, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof.

[0025] In a first embodiment, the present invention relates to a method for treating a CNS disease, health condition, or ailment in an individual in need, comprising administering to the individual, alone or in combination therapy, a therapeutically effective amount of a compound represented by formula I: ; Or a medically acceptable salt thereof, wherein: JC series is selected from the following groups: hydrogen, halogen, C1-6 alkyl, and C1-6 fluoroalkyl with 1 to 3 fluorine atoms substituted; X is either N or C (J C1); J C1 series are selected from the following groups: hydrogen, halogens, C1-6 alkyl groups and C1-6 fluoroalkyl groups substituted with 1 to 3 fluorine atoms; Each JB is independently selected from the group consisting of: hydrogen, halogen, C1-6 alkyl, and C1-6 fluoroalkyl with 1 to 3 fluorine atoms substituted; The JD series is selected from the group consisting of: hydrogen, halogens, C1-6 alkyl groups and C1-6 fluoroalkyl groups substituted with 1 to 3 fluorine atoms; and n is an integer selected from 0, 1, 2, 3 or 4.

[0026] In a second embodiment, the present invention relates to a method for treating a CNS disease, health condition, or ailment in an individual in need, comprising administering, alone or in combination, a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to the individual in a pharmaceutical composition or dosage form.

[0027] In a third embodiment, the present invention further relates to the use of a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the manufacture of an agent for the treatment of CNS diseases, health conditions or ailments of an individual in need.

[0028] In a fourth embodiment, the present invention further relates to a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the treatment of CNS diseases, health conditions or ailments in an individual in need.

[0029] In the first embodiment of the first, second, third and fourth states, for compounds of formula I, n is an integer selected from 1, 2, 3 or 4, each JB is independently selected from the group of the following compositions: halogen, C1-6 alkyl and C1-6 fluoroalkyl substituted with 1 to 3 fluorine atoms, all other carbon atoms of the benzene ring are unsubstituted, and the remaining variables are as defined above.

[0030] In the second embodiment of one of the first, second, third, and fourth state samples, for a compound of formula I or a pharmaceutically acceptable salt thereof, JC is selected from the group consisting of: hydrogen, halogen, and C1-6 alkyl; JCl is selected from the group consisting of: hydrogen, halogen, and C1-6 alkyl; each JB is independently selected from the group consisting of: hydrogen, halogen, and C1-6 alkyl; JD is selected from the group consisting of: hydrogen, halogen, and C1-6 alkyl; and the remaining variables are as defined above for formula I in the first state sample.

[0031] In the third embodiment of one of the first, second, third, and fourth states, the compound of formula I is represented by formula IA: , Or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I of the first state or the first or second embodiment.

[0032] In a fourth embodiment, for a compound of formula IA or a pharmaceutically acceptable salt thereof, JCl is H, F, or Cl; and the remaining variables are as defined in the first state or in the first, second, or third embodiments.

[0033] In a fifth embodiment, JCl is H for a compound of formula IA or a pharmaceutically acceptable salt thereof; and the remaining variables are as defined in the first state or any of the first to fourth embodiments.

[0034] In a sixth embodiment, J C1 is F for compounds of formula IA or their pharmaceutically acceptable salts; and the remaining variables are as defined in the first state or any of the first to fifth embodiments.

[0035] In a seventh embodiment, the compound of formula I is represented by formula IB: , Or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I according to any of the first state sample or the first to sixth embodiments.

[0036] In an eighth embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, n is 2 or 3, and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiments.

[0037] In a ninth embodiment, for compounds of formula I, IA, or IB, or their pharmaceutically acceptable salts, n is 0 or 1, and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiments. In some embodiments, each JB is independently halogenated or C1-6 alkyl.

[0038] In a tenth embodiment, for compounds of formula I, IA, or IB, or their pharmaceutically acceptable salts, each JB is independently H, F, or C1-4 alkyl; and the remaining variables are as described in the first embodiment or in the first, second, third, fourth, fifth, sixth, seventh, eighth, or ninth embodiments. In some embodiments, each JB is independently F or C1-4 alkyl.

[0039] In an eleventh embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, n is 2 or 3; each JB is independently F or methyl; and the remaining variables are as described in the first state or in the first, second, third, fourth, or fifth, sixth, or seventh embodiments.

[0040] In a twelfth embodiment, for compounds of formula I, IA, or IB, or their pharmaceutically acceptable salts, n is 2; JB are all F or one of JB is F and the other is methyl; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiment. In some embodiments, one JB is F and the other is methyl.

[0041] In a thirteenth embodiment, for compounds of formula I, IA, or IB, or their pharmaceutically acceptable salts, n is 3; two of JB are F and the other is methyl; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiment. In some embodiments, JD is H or F. In some embodiments, JD is F. In some embodiments, JD is H.

[0042] In a fourteenth embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, n is 1; JB is F; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiment.

[0043] In a fifteenth embodiment, n is 0 for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, or seventh embodiment.

[0044] In a sixteenth embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, JD is hydrogen; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, or fifteenth embodiments.

[0045] In a seventeenth embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, JD is F, Cl, or methyl; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, or fifteenth embodiments.

[0046] In an eighteenth embodiment, JD is F for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, or fifteenth embodiments.

[0047] In a nineteenth embodiment, for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts, JC is H, Cl, or F; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, or eighteenth embodiments.

[0048] In a twentieth embodiment, JC is H for compounds of formula I, IA, or IB or their pharmaceutically acceptable salts; and the remaining variables are as described in the first state or in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, or eighteenth embodiments.

[0049] In a twenty-first embodiment, the compound of formula I is a compound shown in Table I or a pharmaceutically acceptable salt thereof. [surface] [I.] [illustrative] [sGC] [Stimulants.] Or a medically acceptable salt.

[0050] In a twenty-second embodiment, for the method and use of the present invention, the sGC stimulant is compound I-14 or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutically acceptable salt is a sodium salt. In another embodiment, the sGC stimulant is a sodium salt of compound I-14 represented by the following formula: .

[0051] In a twenty-third embodiment, for the method and use of the present invention, the sGC stimulant is compound I-20 or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutically acceptable salt is a sodium salt. In another embodiment, the sGC stimulant is a sodium salt of compound I-20 represented by the following formula: .

[0052] In a twenty-fourth embodiment, the compound of the present invention is a compound represented by formula IC: , Or a pharmaceutically acceptable salt thereof, wherein X is N or C (J C1), wherein when X is C (J C1), it is represented by C in the table below; and the definitions of the variables X, J C1, and JB are described in the table below; furthermore, Me represents methyl and Me-F represents a fluorinated methyl group substituted with 1 to 3 fluorine atoms (i.e., -CH2F, -CHF2, or CF3): [Compound Number] [J, B , ] [At] [Position on the benzene ring and J, B , ] [Definition] [X] [J, C1 , ] [2] [3] [4] [5] [6] I - 27 F C F I - 28 F C H I - 29 F N - I - 30 F C F I - 31 F F C F I - 32 F F C H I-33 F F N - I-34 F F C F I-35 F F C H I-36 F F N - I-37 F F C F I-38 F F C H I-39 F F N - I-40 F F C F I-41 F F C F I-42 F F C H I-43 F F N - I-44 F F C F I-45 F Me C H I-46 F Me N - I-47 F Me C F I-48 Me F C H I-49 Me F N - I-50 Me F C F I-51 F Me C H I-52 F Me N - I-53 F Me C F I-54 Me F C H I-55 Me F N - I-56 Me F C F I-57 F Me C H I-58 F Me C F I-59 F Me N - I-60 Me F C H I-61 Me F C F I-62 Me F N - I-63 F Me C H I-64 F Me N - I-65 F Me C F I-66 Me F C H I-67 Me F N - I-68 Me F C F I-69 F Me N - I-70 F Me C F I-71 Me F C H I-72 Me F N - I-73 Me F C F I-74 F Me C H I-75 F Me N - I-76 F Me C F I-77 Me F C H I-78 Me F N - I-79 Me F C F I-80 Me F C H I-81 Me F N - I-82 Me F C F I-83 F Me C H I-84 F Me N - I-85 F Me C F I-86 F F Me C H I-87 F F Me N - I-88 F F Me C F I-89 F F Me C H I-90 F F Me N - I-91 F F Me C F I-92 F F Me C H I-93 F F Me N - I-94 F F Me C F I-95 F Me F C H I-96 F Me F N - I-97 F Me F C F I-98 F F Me C H I-99 F F Me N - I-100 F F Me C F I-101 F F Me C H I-102 F F Me N - I-103 F F Me C F I-104 F Me F C H I-105 F Me F C F I-106 F Me F N - I-107 F F Me C H I-108 F F Me C F I-109 F F Me N - I-110 F Me F C H I-111 F Me F N - I-112 F Me F C F I-113 F Me F C H I-114 F Me F N - I-115 F Me F C F I-116 Me F F C H I-117 Me F F N - I-118 Me F F C F I-119 F F Me C H I-120 F F Me N - I-121 F F Me C F I-122 F F Me C H I-123 F F Me N - I-124 F F Me C F I-125 Me F F C H I-126 Me F F N - I-127 Me F F C F I-128 F Me F N - I-129 F Me F C F I-130 F Me-F C H I-131 F Me-F N - I-132 F Me-F C F I-133 Me-F F C H I-134 Me-F F N - I-135 Me-F F C F I-136 F Me-F C H I-137 F Me-F N - I-138 F Me-F C F I-139 Me-F F C H I-140 Me-F F N - I-141 Me-F F C F I-142 F Me-F C H I-143 F Me-F C F I-144 F Me-F N - I-145 Me-F F C H I-146 Me-F F C F I-147 Me-F F N - I-148 F Me-F C H I-149 F Me-F N - I-150 F Me-F C F I-151 Me-F F C H I-152 Me-F F N - I-153 Me-F F C F I-154 F Me-F N - I-155 F Me-F C F I-156 F Me-F C H I-157 Me-F F N - I-158 Me-F F C H I-159 Me-F F C F I-160 F Me-F C H I-161 F Me-F N - I-162 F Me-F C F I-163 Me-F F C H I-164 Me-F F N - I-165 Me-F F C F I-166 Me-F F C H I-167 Me-F F N - I-168 Me-F F C F I-169 F Me-F C H I-170 F Me-F N - I-171 F Me-F C F I-172 F F Me-F C H I-173 F F Me-F N - I-174 F F Me-F C F I-175 F F Me-F C H I-176 F F Me-F N - I-177 F F Me-F C F I-178 F F Me-F C H I-179 F F Me-F N - I-180 F F Me-F C F I-181 F Me-F F C H I-182 F Me-F F N - I-183 F Me-F F C F I-184 F F Me-F C H I-185 F F Me-F N - I-186 F F Me-F C F I-187 F F Me-F C H I-188 F F Me-F N - I-189 F F Me-F C F I-190 F Me-F F C H I-191 F Me-F F C F I-192 F Me-F F N - I-193 F F Me-F C H I-194 F F Me-F C F I-195 F F Me-F N - I-196 F Me-F F C H I-197 F Me-F F N - I-198 F Me-F F C F I-199 F Me-F F C H I-200 F Me-F F N - I-201 F Me-F F C F I-202 Me-F F F C H I-203 Me-F F F N - I-204 Me-F F F C F I-205 F F Me-F C H I-206 F F Me-F N - I-207 F F Me-F C F I-208 F F Me-F C H I-209 F F Me-F N - I-210 F F Me-F C F I-211 Me-F F F C H I-212 Me-F F F N - I-213 Me-F F F C F I-214 F Me-F F N - I-215 F Me-F F C F I-216 F Me-F F C H

[0053] [, , ] [, The present invention relates to a medically acceptable salt. , ] [, , ] The term "pharmaceutically acceptable salt" as used herein includes salts of compounds obtained by mixing such compounds with inorganic or organic acids or bases. In some embodiments, such salts may be prepared in situ during the final separation and purification of the compound. In other embodiments, salts may be prepared from the free form of the compound during individual synthetic steps. The preparation of pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts is more fully described in Berg et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977:66:1-19, which is incorporated herein by reference in its entirety. Pharmaceutically acceptable salts of Formula I compounds are those salts that can be used in pharmaceuticals. However, pharmaceutically unacceptable salts may be used to prepare compounds of Formula I or their pharmaceutically acceptable salts.

[0054] When the compound of Formula I is acidic, a suitable "medically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable, non-toxic base, including both inorganic and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, trivalent manganese salts, divalent manganese salts, potassium salts, sodium salts, zinc salts, and similar salts. Specific examples include ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts. Salts derived from pharmaceutically acceptable organic nontoxic bases include salts of primary, secondary, and tertiary amines, including naturally occurring substituted amines, cyclic amines, arginine, betaine, caffeine, choline, N,N-1-diphenylmethylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylpyridine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, hyprothidine, isopropylamine, lysine, methylglucosamine, pyridine, piperidine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, thiazolinone, and their analogues.

[0055] In some embodiments, the compound of formula I has an acidic OH group that can react with a base (e.g., a pharmaceutically acceptable non-toxic base) to form a salt (e.g., a pharmaceutically acceptable salt). In some embodiments, the salt is an ammonium salt, calcium salt, magnesium salt, potassium salt, or sodium salt. In other embodiments, the salt is a sodium salt.

[0056] When the compound of formula I is a base, the salt can be prepared from pharmaceutically acceptable, non-toxic acids (including inorganic and organic acids). Such acids include acetate, acetic acid, acidic citrate, acidic phosphate, ascorbate, benzenesulfonic acid, benzenesulfonate, benzoic acid, benzoate, bromide, hydrogen sulfate, hydrogen tartrate, camphorsulfonic acid, chloride, citrate, citric acid, ethanesulfonate, ethanesulfonic acid, formate, fumarate, fumaric acid, gentianate, gluconic acid, glucuronic acid, glutamate, glutamic acid, hydrobromic acid, hydrochloric acid, iodide, hydroxyethylsulfonic acid, isonicotinic acid, and lactic acid. Radicals, lactic acid, maleic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, methanesulfonate, mucilage, nitrate, nitric acid, oleate, oxalate, pyruvic acid, bis(hydroxynaphthyl)ate (i.e., 1,1'-methylene-bis(2-hydroxy-3-naphthylcarboxylate)), pantothenic acid, pantothenate, phosphate, phosphoric acid, glucoside, salicylate, succinic acid, succinic acid, sulfuric acid, sulfate, tannic acid, tartrate, tartaric acid, p-toluenesulfonate, p-toluenesulfonic acid, and the like. Specific embodiments include citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid. [, , ]

[0057] In addition to the compounds described herein, pharmaceutically acceptable salts of these compounds may also be used in compositions or dosage forms to treat or prevent the diseases identified herein.

[0058] [, Pharmaceutical compositions, dosage forms, and administration methods. , ] [, , ] The compounds disclosed herein and their pharmaceutically acceptable salts may be formulated into pharmaceutical compositions or "formulations" for the therapeutic and therapeutic uses of the present invention.

[0059] Typical formulations are prepared by mixing a compound of Formula I or a pharmaceutically acceptable salt thereof with a carrier, diluent, or excipient. Suitable carriers, diluents, and excipients are well known to those skilled in the art and include substances such as carbohydrates, waxes, water-soluble and / or expandable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, water, and the like. The specific carrier, diluent, or excipient used will depend on the means and purpose of formulating the compound of Formula I. Solvents are generally selected based on those skilled in the art who recognize them as generally safe for mammals (GRAS). Generally, safe solvents are non-toxic aqueous solvents, such as water and other non-toxic solvents that are soluble or miscible with water. Suitable aqueous solvents include water, ethanol, propylene glycol, polyethylene glycol (e.g., PEG400, PEG300), and mixtures thereof. The formulation may also include other types of excipients, such as one or more buffers, stabilizers, anti-adhesives, surfactants, humectants, lubricants, emulsifiers, binders, suspending agents, disintegrants, fillers, adsorbents, coatings (e.g., enteric or slow-release), preservatives, antioxidants, light-blocking agents, flow aids, processing aids, colorants, sweeteners, flavorings, and other known additives used to provide a refined presentation of the medicine (i.e., a compound of formula I or a pharmaceutical composition thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a pharmaceutical preparation).

[0060] Acceptable diluents, carriers, excipients, and stabilizers are substances that are non-toxic to the recipient at the doses and concentrations used, and include buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethylbenzyl ammonium chloride; hexahydroxyquaternary ammonium chloride; benzalkonium chloride, benzyl ammonium chloride; phenol, butanol, or benzyl alcohol; alkyl parabens such as methylparaben or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-methylparaben). Phenols); proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamic acid, aspartic acid, histamine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; salt-forming ions, such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG). Active pharmaceutical ingredients can also be encapsulated in prepared microcapsules, for example, using coagulation techniques or interfacial polymerization. Examples include hydroxymethyl cellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules encapsulated in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's: The Science and Practice of Pharmacy, 21st edition, University of the Sciences in Philadelphia, 2005 (hereinafter referred to as "Remington's").

[0061] The formulation can be prepared using known dissolving and mixing procedures.

[0062] As used herein, the term "therapeutic effective amount" means the amount of an active compound or pharmaceutical agent that elicits a biological or pharmaceutical response sought by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, or human. The therapeutic effective amount of a compound to be administered will be determined by such considerations and is the minimum amount required to improve, cure, or treat one or more of the disease or its symptoms.

[0063] The term "administer / administering / administration" in relation to the compounds, compositions, or dosage forms of this invention means the introduction of the compound into the system of an individual or a patient requiring treatment. When the compounds of this invention are provided in combination with one or more other active agents, "administer" and its variations are each understood to include the simultaneous and / or sequential introduction of the compound and other active agents.

[0064] Depending on the severity and type of the disease to be treated, the compositions described herein may be administered systemically or topically, such as orally (including, but not limited to, solid dosage forms, including hard or soft capsules (e.g., gelatin capsules), tablets, pills, powders, sublingual tablets, sugar-coated tablets, lozenges, and granules); and liquid dosage forms, including, but not limited to, pharmaceutically acceptable emulsions, microemulsions, aqueous or oily solutions, suspensions, syrups, and elixirs); by inhalation (e.g., using aerosols, gases, inhalers, nebulizers, or the like); by ear (e.g., using ear drops); and by topical application (e.g., using creams, gels, inhalers, liniments, lotions, ointments, etc.). Intra-abortive methods include: patches, pastes, powders, solutions, sprays, percutaneous patches, etc.; ocular (e.g., using eye drops, ocular gels, ocular ointments); rectal (e.g., using enemas or suppositories); nasal; buccal; vaginal (e.g., using irrigators, intrauterine devices, vaginal suppositories, vaginal rings or lozenges, etc.); ear drops; implantable reservoirs or the like; or non-intestinal. As used herein, the term "non-intestinal" includes (but is not limited to) subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally, or intravenously.

[0065] Formulations of compounds intended for oral use can be prepared according to any method known in this art for manufacturing pharmaceutical compositions.

[0066] In solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silica; b) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; c) humectants, such as glycerin; d) disintegrants, such as agar-agar, calcium carbonate, potato or cassava starch, alginic acid, certain silicates, and sodium carbonate; e) solvent inhibitors, such as paraffin; f) absorption enhancers, such as quaternary ammonium compounds; g) humectants, such as cetyl alcohol and glyceryl monostearate; h) absorbents, such as kaolin and bentonite; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. The tablets may be uncoated or coated using known techniques, including microencapsulation, to mask unpleasant tastes or to delay disintegration and absorption in the gastrointestinal tract and / or to provide sustained action over a longer period. For example, delaying agents such as glyceryl monostearate or glyceryl distearate may be used alone or in combination with waxes. Water-soluble taste-masking materials such as hydroxypropyl-methylcellulose or hydroxypropyl-cellulose may be used.

[0067] In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in this technology, such as water or other solvents; solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, methyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, wheat germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol; and mixtures thereof. Besides inert diluents, oral compositions may also include adjuvants, such as humectants, emulsifiers and suspending agents, sweeteners, flavoring agents, and aromatizers.

[0068] Oral compositions (solid or liquid) may also include excipients and adjuvants, such as dispersants or humectants, such as naturally occurring phospholipids (e.g., lecithin); condensation products of alkyl esters and fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide and long-chain aliphatic alcohols (e.g., heptadecanethoxylated hexadecyl alcohol), condensation products of ethylene oxide and partial esters derived from fatty acids and hexadiol anhydrides (e.g., polyoxyethylene sorbitan monooleate); emulsifiers and suspending agents, such as sodium carboxymethyl cellulose, croscarmellose, povidone, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, gum arabic, and gum arabic; sweeteners, flavoring agents, and aroma agents; and / or one or more preservatives, such as ethylparaben or n-propylparaben; one or more colorants; one or more flavoring agents; and one or more sweeteners, such as sucrose or saccharin.

[0069] Pharmaceutical compositions can also be administered via nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in pharmaceutical formulation and can be prepared as a solution in physiological saline using benzyl alcohol or other suitable preservatives, absorption enhancers to improve bioavailability, fluorocarbons, and / or other known solvents or dispersants. Suitable for intrapulmonary or nasal administration, the particle size of the formulation is, for example, in the range of 0.1 micrometers to 500 micrometers (including particles in the range of 0.1 micrometers to 500 micrometers increasing in micrometers such as 0.5 micrometers, 1 micrometer, 30 micrometers, 35 micrometers, etc.), delivered to the alveolar sacs by rapid inhalation through the nasal cavity or by oral inhalation.

[0070] The pharmaceutical compositions described herein can also be administered topically, especially when the therapeutic target includes areas or organs easily accessible by topical application (including diseases of the eyes, ears, skin, or lower intestine). Suitable topical formulations for each of these areas or organs can be readily prepared. The active ingredient is blended under sterile conditions with a pharmaceutically acceptable carrier and, if applicable, any desired preservatives or buffers.

[0071] For topical application, pharmaceutical compositions may be formulated in a suitable ointment form containing an active ingredient suspended or dissolved in one or more carriers. Carriers used for topical application of the compounds of the present invention include (but are not limited to) mineral oils, liquid paraffins, white paraffins, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsified waxes, and water. Alternatively, pharmaceutical compositions may be formulated in a suitable emulsion or cream form containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oils, sorbitan monostearate, polysorbate 60, hexadecyl ester wax, cetearyl alcohol, 2-octyldodecyl alcohol, benzyl alcohol, and water.

[0072] Alternatively, the active ingredient may be formulated into a cream together with an oil-in-water emulsion base. Where necessary, the aqueous phase of the emulsion base may include polyols, i.e., alcohols having two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol, and polyethylene glycol (including PEG 400) and mixtures thereof.

[0073] Surface formulations may include compounds that enhance the absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such transdermal penetration enhancers include dimethyl sulfoxide and similar compounds.

[0074] The oil phase of an emulsion prepared using the compounds in Table I may be composed of known components in a known manner. Although this phase may contain only emulsifiers (or laxatives), it preferably contains at least one emulsifier with fats or oils, or with a mixture of both fats and oils. Hydrophilic emulsifiers may be included together with lipophilic emulsifiers that act as stabilizers. In some embodiments, the emulsifier includes both oils and fats. The emulsifier, together with or without the stabilizer, constitutes a so-called emulsified wax, and the wax, together with the oils and fats, constitutes a so-called emulsified ointment base, which forms the oily dispersed phase of the cream formulation. Laxatives and emulsion stabilizers suitable for formulations of compounds of Formula I include Tween™-60, Span™-80, cetearyl alcohol, benzyl alcohol, myristyl alcohol, glyceryl monostearate, and sodium lauryl sulfate.

[0075] Furthermore, this invention covers the use of transdermal patches, which offer the added advantage of controlled delivery of compounds to the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the transdermal amount of the compound. The rate can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0076] For ocular use, the pharmaceutical composition may be formulated as a micron-sized suspension in isotonic pH-adjusted sterile saline, or preferably as a solution in isotonic pH-adjusted sterile saline, with or without a preservative (such as benzyl chlorobenzyl ammonium). Alternatively, for ocular use, the pharmaceutical composition may be formulated as an ointment (such as paraffin oil). For treatment of the eyes or other external tissues (e.g., mouth and skin), the formulation may be applied as a topical ointment or cream containing, for example, 0.075 to 20% w / w of the active ingredient. When formulated as an ointment, the active ingredient may be used with an oil-based paraffin or a water-miscible ointment base.

[0077] For transrectal or transvaginal administration, the composition is preferably in the form of a suppository, which can be prepared by mixing the compound described herein with a suitable non-irritating excipient or carrier (such as cocoa butter, beeswax, polyethylene glycol, or suppository wax, which is solid at ambient temperature but liquid at body temperature, and thus melts and releases the active compound in the rectal or vaginal cavity). Other formulations suitable for transvaginal administration may be presented in the form of pessaries, tampons, creams, gels, ointments, foams, or sprays.

[0078] The sterile injectable forms of the compositions described herein (e.g., for non-enteric administration) may be aqueous or oily suspensions. These suspensions may be formulated using suitable dispersants or wetting agents and suspending agents (including those described in the preceding paragraphs) according to techniques known in this art. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic, non-enteric acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Among these acceptable mediators and solvents, water, Ringer's solution, and isotonic sodium chloride solution may be used. Additionally, sterile non-volatile oils are commonly used as solvents or suspension media. For this purpose, any mild non-volatile oil, including synthetic monoglycerides or diglycerides, may be used. Fatty acids (such as oleic acid and its glycerol derivatives) are suitable for the preparation of injectable formulations in the form of naturally pharmaceutically acceptable oils (such as vegetable oils, e.g., peanut oil, olive oil, sesame oil, or coconut oil), particularly in their polyoxyethylene form or in mineral oils such as liquid paraffin. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethyl cellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms (including emulsions and suspensions). Other commonly used surfactants (such as Tween, Span, and other emulsifiers) or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for injectable formulations. Oily suspensions may contain thickeners such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners (such as those mentioned above) and flavoring agents may be added to provide palatable oral formulations. These compositions may be preserved by adding antioxidants such as butylated hydroxyanisole or α-tocopherol.

[0079] In another embodiment, a compound of formula I or a pharmaceutically acceptable salt thereof may be formulated into a veterinary composition comprising a veterinary carrier. The veterinary carrier is a material suitable for administration of the composition and may be a solid, liquid, or otherwise inert gaseous material. It is compatible with the active ingredient in the veterinary field. These veterinary compositions may be administered via non-intestinal, oral, or any other desired route of administration.

[0080] [, Treatment , ] In one first embodiment, the present invention relates to a method for treating a CNS disease, health condition, or ailment in an individual in need, comprising administering to the individual, alone or in combination therapy, a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of formula I is as described in any of the first to twenty-first embodiments above.

[0081] In a second embodiment, the present invention relates to a method for treating a CNS disease, health condition, or ailment in an individual in need, comprising administering, alone or in combination therapy, a therapeutically effective amount of a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof to the individual. In some embodiments, the compound of formula I is as described in any of the first to twenty-first embodiments above.

[0082] In a third embodiment, the invention further relates to the use of a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the manufacture of an agent for treating a CNS disease, health condition, or ailment of an individual in need. In some embodiments, the compound of formula I is as described in any of the first to twenty-first embodiments above.

[0083] In a fourth embodiment, the invention further relates to a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form comprising a compound of formula I or a pharmaceutically acceptable salt thereof, for the treatment of CNS diseases, health conditions, or symptoms in an individual in need. In some embodiments, the compound of formula I is as described in any of the first to twenty-first embodiments above.

[0084] In one embodiment of the first, second, third, and fourth embodiments, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by increased neuroinflammation. One embodiment of the invention is a method for reducing neuroinflammation in an individual by administering to that individual a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing thereof.

[0085] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by increased neurotoxicity. One embodiment of the present invention is a method for reducing or compensating for the negative neurotoxicity of an individual by administering to an individual any of a compound of formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition or dosage form containing thereof.

[0086] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by impaired neuroregeneration. One embodiment of the present invention is a method for restoring neuroregeneration in an individual by administering any of the following to an individual in need: a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing the same.

[0087] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by weakened synaptic function. One embodiment of the present invention is a method for restoring synaptic function in an individual by administering to that individual any of a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing thereof.

[0088] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by downregulated neurotransmitters. One embodiment of the present invention is a method for normalizing the neurotransmitters of an individual by administering to that individual any of a compound of formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition or dosage form containing thereof.

[0089] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by reduced cerebral blood flow. One embodiment of the present invention is a method for restoring cerebral blood flow to an individual by administering any of the following to an individual in need: a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing the same.

[0090] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by increased neurodegeneration. One embodiment of the present invention is a method for reducing neurodegeneration in an individual by administering any of the following to an individual in need: a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing such a compound.

[0091] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of diseases and conditions characterized by cognitive impairment. One embodiment of the present invention is a method for improving the cognition of an individual by administering to an individual any of a compound of formula I or a pharmaceutically acceptable salt thereof or a pharmaceutical composition or dosage form containing thereof. In some embodiments, the treatment improves memory. In other embodiments, the treatment improves attention. In other embodiments, the treatment improves executive function.

[0092] In another embodiment of the first to fourth embodiments of the present invention, the compounds disclosed herein are neuroprotective sGC stimulants. Specifically, compounds of formula I or their pharmaceutically acceptable salts or pharmaceutical compositions or dosage forms containing them may be suitable for protecting neurons in individuals in need.

[0093] In some embodiments, CNS diseases, health conditions, or disorders are selected from Alzheimer's disease (AD), vascular dementia (VD), vascular cognitive impairment, mixed dementia, Binswanger's dementia (subcortical arteriosclerosis encephalopathy), CADASIL or CADASIL syndrome with subcortical infarction and leukoencephalopathy, frontotemporal lobe degeneration or dementia (FTD), asymptomatic neurocognitive impairment (ANI), subjective cognitive impairment or decline (SCD), cognitive aging, minimal neurocognitive disorder (MND), HIV-associated dementia (HAD) (also known as AIDS dementia complex [ADC] or HIV encephalopathy), Lewy body dementia, Alzheimer's disease, or mild cognitive impairment (MCI).

[0094] In other embodiments, the disease, health condition, or symptom is selected from the following CNS disorders or symptoms: sleep-wake disorders and neurological abnormalities associated with Sneddon syndrome.

[0095] In other embodiments, the disease, health condition, or condition is a CNS condition or condition selected from the group consisting of: Alzheimer's disease or early Alzheimer's disease, mild to moderate Alzheimer's disease, or moderate to severe Alzheimer's disease.

[0096] In other embodiments, the CNS disease is selected from the group consisting of glaucoma, Huntington's disease (or Huntington's chorea chorea), HD), multiple sclerosis (MS), multiple system dystrophy (MSA), Parkinson’s disease (PD), Parkinson’s superposition syndrome, spinocerebellar ataxia (SCA), Steel-Richardson-Olszewski disease (progressive supranuclear palsy), muscular dystrophic lateral sclerosis (ALS or Lou Gehrig’s disease) or ou Gehrig’s disease syndrome).

[0097] In other embodiments, the CNS disease is selected from the group consisting of attention deficit disorder (ADD) and attention deficit hyperactivity disorder (ADHD).

[0098] In other embodiments CNS conditions are traumatic (closed or open) head puncture injury, traumatic brain injury (TBI), nontraumatic stroke (specifically, ischemic stroke), aneurysm, hypoxia, or other damage to the brain.

[0099] In other embodiments, the CNS condition is a mental, psychological, emotional, or affective condition selected from the group consisting of: bipolar disorder, schizophrenia, general psychosis, drug-induced psychosis, delusional disorder, schizoaffective disorder, obsessive-compulsive disorder (OCD), depression, anxiety disorder, panic disorder, or post-traumatic stress disorder (PTSD).

[0100] In other embodiments, the CNS disease or condition is selected from muscle hypotonia, including, for example, generalized, local, segmental, sexual, intermediate, hereditary / primary muscle dystonia or acute dystonic reaction (acute dystonic reaction);

[0101] In other embodiments, CNS diseases or conditions are selected from conditions characterized by a relative reduction in synaptic plasticity and synaptic processes, including, for example, Fragile X, Rhett's disorder, Williams syndrome, Renpenning syndrome, ASD, autism, Asperger's syndrome, pervasive developmental disorder, or childhood breakdown disorder.

[0102] In other embodiments, CNS symptoms are selected from chemical brain, levodopa-induced addictive behavior, alcohol poisoning, narcotic dependence (including but not limited to amphetamine, opiates or other substances), and drug abuse.

[0103] In one embodiment, a CNS disease is a cognitive or functional impairment caused by brain injury, mental illness, neurodevelopmental disorder, or neurodegenerative disorder.

[0104] In some embodiments of the methods and uses of the present invention, cognitive impairment (such as MCI or dementia) is associated with the following: Alzheimer's disease (AD), vascular dementia, mixed dementia, AD with vascular lesions (ADv), cerebral infarction, cerebral ischemia, stroke, head injury, traumatic head injury, learning disability, autism, attention deficit disorder, depression, spinocerebellar ataxia, Lewy body dementia, dementia with frontal lobe degeneration, Pick's syndrome, Parkinson's disease, progressive nuclear palsy, dementia with corticobasal ganglia degeneration, amyotrophic lateral sclerosis (ALS), Huntington's disease, demyelination, multiple sclerosis (MS), thalamic degeneration, Creutzfeldt-Jakob dementia, HIV-related dementia, schizophrenia, and Korsakoff's psychosis. (psychosis), postoperative cognitive decline in older adults, bipolar disorder, or mitochondrial disorders. In other embodiments, cognitive impairment is associated with sickle cell disease.

[0105] In some embodiments of the methods and uses of the present invention, MCI, dementia, subclinical cognitive impairment, or SCD are associated with: cognitive aging, postoperative cognitive decline, drug side effects, metabolic imbalance, hormonal problems, vitamin or nutrient deficiencies, delirium, mental illness, damage to brain neurons due to an injury (e.g., stroke or other cerebrovascular disease or due to traumatic brain injury), early neurodegenerative processes, exposure to toxins, or viral or bacterial infections.

[0106] In other embodiments, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of orphan pain indications. One embodiment of the invention is a method for treating orphan pain indications by administering to an individual in need any of the following: a compound of formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or dosage form containing the compound. Specifically, the orphan pain indication is selected from acetazolamide-responsive myotonia, autoimmune syndrome, and autosomal dominant Charcot-Marie-Tooth disease type 2V. 2V), autosomal dominant moderate Chuck-Maley-Duss disease with neuralgia, autosomal recessive limb-girdle muscular dystrophy type 2A, congenital analgesia associated with ion channelopathies, chronic pain requiring intraspinal analgesia, complex regional pain syndrome, complex regional pain syndrome type 1, complex regional pain syndrome type 2, congenital analgesia with hyperhidrosis, congenital analgesia with severe intellectual disability, congenital analgesia with hypohidrosis, diffuse palmar erythema with painful fissures. Plantar keratoderma, familial intermittent pain syndrome, familial intermittent pain syndrome with significant lower limb involvement, familial intermittent pain syndrome with major upper body involvement, hereditary painful calluses, hereditary sensory and autonomic lesions type 4, hereditary sensory and autonomic lesions type 5, hereditary sensory and autonomic lesions type 7, interstitial cystitis, painful orbital and generalized neurofibromatosis-Marfanoid habitus syndrome, paroxysmal severe pain, persistent idiopathic facial pain, qualitative or quantitative calpain deficiency, Tolosa-Hunt syndrome.

[0107] In other embodiments, the CNS symptom is neuropathic pain. In some embodiments, the pain is neuropathic pain associated with a CNS disease.

[0108] In other embodiments, the disease or symptom is selected from acute pain, central pain syndrome, chemotherapy-induced neuropathy, diabetic neuropathy, myofibromyalgia, inflammatory pain, painful diabetic peripheral neuropathy, postoperative pain, ankylosing spondylitis, and visceral pain.

[0109] In other embodiments, the compounds disclosed herein are sGC stimulants suitable for the prevention and / or treatment of the following: high altitude (altitude) disease, cerebral small vessel disease, cerebral vasculitis, cerebral vasospasm, hepatic encephalopathy, moyamoya disease, Parkinson's disease, dysphagia, ataxia, telangiectasia, autism spectrum disorder, chronic fatigue, chronic traumatic encephalopathy (CTE), cognitive impairment associated with diabetes, cognitive impairment associated with multiple sclerosis, cognitive impairment associated with obstructive sleep apnea, cognitive impairment associated with schizophrenia (CIAS), cognitive impairment associated with sickle cell disease, concussion, retinopathy, diabetic retinopathy (including proliferative and non-proliferative), and dysphagia.

[0110] In other embodiments, the compounds disclosed herein are suitable for the prevention and / or treatment of the following sGC stimulants: ocular fibrosis, Fabry disease, Gaucher disease, glioblastoma, encephalitis symptomatic of cerebral malaria (SoC), encephalitis caused by infectious diseases, intellectual disability, myopic choroidal neovascularization, neuromyelitis optica, neuralgia with multiple sclerosis, neuralgia with shingles / herpes zoster, neuralgia following spinal surgery, Parkinson's disease, peripheral and autonomic neuropathy, peripheral retinal degeneration, post-traumatic stress disorder, postherpetic neuralgia, postoperative dementia, proliferative vitreoretinopathy, radiation-induced cerebral fibrosis, radiculopathy, refractory epilepsy, retinal vein occlusion, spinal cord injury, spinal muscular atrophy, spinal subluxation, tau proteinosis, and wet age-related macular degeneration.

[0111] The CNS diseases that can benefit from treatment with the sGC stimulant of the present invention are those in which an increase in NO concentration or an increase in cGMP concentration or both, or an upregulation of the NO-sGC-cGMP pathway, is appropriate.

[0112] The compounds described herein, as well as their pharmaceutically acceptable salts, are sGC stimulants capable of crossing the blood-brain barrier (BBB) ​​and are suitable for the prevention and / or treatment of CNS diseases, symptoms, and conditions that may benefit from sGC stimulation in the brain.

[0113] In some embodiments, the compounds of the present invention can stimulate sGC in the brain without causing a significant decrease in blood pressure (BP) in the patient. In some embodiments, the compounds, at doses that induce the desired CNS effect, cause an average decrease in BP of less than 5 mm Hg in the patient. In some embodiments, the average decrease is less than 10 mm Hg. In other embodiments, the decrease in BP in the patient is not clinically significant. In still other embodiments, the methods and uses of the present invention do not cause a significant incidence of adverse events (AEs) associated with symptomatic hypotension when treating patients with CNS disorders.

[0114] As used herein, a dose that does not "cause a significant incidence of adverse events (AEs) associated with symptomatic hypotension" is a dose that does not cause excessive orthostatic hypotension, excessive vertigo, excessive orthostatic vertigo, excessive presyncope, or excessive syncope in patients. Excessive orthostatic hypotension, excessive vertigo, excessive orthostatic vertigo, excessive presyncope, or excessive syncope in patients are those circumstances that warrant the discontinuation of treatment or the discontinuation of recommendations by the physician.

[0115] In some embodiments, BP is measured as systolic BP. In other embodiments, diastolic BP is measured. In other embodiments, mean arterial pressure (MAP) is measured. In other embodiments, BP is measured while walking. In other embodiments, BP is measured at the wrist. In some embodiments, BP is measured with the patient in a supine position. In other embodiments, the patient is in a sitting position.

[0116] As used herein, the term "disease" means any deviation or disruption of the normal structure or function of any part, organ, or system of the body, characterized by a set of symptoms and signs, the cause, pathology, and prognosis of which may be known or unknown. The term "disease" encompasses other related terms such as symptoms and conditions (or medical conditions) and syndromes, defined as a combination of symptoms that are caused by or consequently relate to a single cause and typically occur together when forming different clinical pictures. In some embodiments, the term "disease" refers to a medical or pathological disease mediated by sGC, cGMP, and / or NO. In some embodiments, the term "disease" refers to a medical or pathological disease mediated by sGC, cGMP, and / or NO affecting the CNS. As used herein, "CNS disease, health condition, or symptom" means the same as CNS disease, CNS health condition, or CNS symptom, and the terms disease, symptom, and symptom in this invention, regardless of their usage, refer only to those affecting the CNS (central nervous system).

[0117] "Treatment" in relation to a disease, condition, symptom, or syndrome means eliminating or improving the cause and / or effects of that disease, condition, or syndrome (i.e., symptoms, physiological, physical, mental, emotional, or any other clinical manifestation, observation, or measurement, or improvement of pathological assessment).

[0118] As used in this article, the term “treatment” also means delaying or improving or preventing the progression of disease (i.e., known or anticipated progression of disease), its severity and / or duration, or delaying or improving or preventing the progression of one or more symptoms, clinical manifestations, observations or measurements, or preventing or mitigating the negative developmental progression of a pathological assessment caused by the administration of one or more therapies (i.e., “controlling” rather than “curing” the condition).

[0119] As used herein, the terms "individual" and "patient" are used interchangeably. The terms "individual" and "patient" refer to animals (e.g., birds such as chickens, quails, or turkeys, or mammals), particularly including non-primates (e.g., cattle, pigs, horses, sheep, rabbits, guinea pigs, rats, cats, dogs, and mice) and primates (e.g., monkeys, chimpanzees, and humans), and more specifically, humans. In some embodiments, an individual is a non-human animal, such as a farm animal (e.g., a horse, cattle, pig, or sheep) or a companion animal or pet (e.g., a dog, cat, mouse, rat, hamster, gerbil, guinea pig, or rabbit). In some embodiments, an individual is a human.

[0120] The present invention also provides a method for treating one of the above-mentioned diseases in an individual, comprising administering to the individual requiring treatment a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof. Alternatively, the present invention provides the use of a compound of formula I or a pharmaceutically acceptable salt thereof for treating one of the diseases in an individual requiring treatment. The present invention also includes the use of a compound of formula I or a pharmaceutically acceptable salt thereof for manufacturing an agent for treating one of the above-mentioned diseases in an individual requiring treatment. The present invention further provides a method for manufacturing an agent suitable for treating one of the diseases, comprising using a compound of formula I or a pharmaceutically acceptable salt thereof.

[0121] As used herein, the term "biological sample" means a live or ex vivo sample, including (but not limited to) cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; blood, saliva, urine, excrement, semen, tears, lymph, intraocular fluid, vitreous fluid, cerebrospinal fluid (CSF) or other bodily fluids or extracts thereof.

[0122] In other embodiments, the present invention provides a method for stimulating sGC activity in a biological sample, comprising contacting the biological sample with a compound or composition of the present invention. The use of sGC stimulants in biological samples is suitable for a variety of purposes known to those skilled in the art. Examples of such purposes include (but are not limited to) bioanalysis and biosample storage.

[0123] [, Combination therapy , ] [, , ] The compounds and pharmaceutical compositions described herein may be used alone or in combination with therapies for the treatment of diseases mediated, regulated or affected by sGC, cGMP and / or NO.

[0124] As used herein, the terms "combination" (as in the phrase "combination therapy") or "co-administered" are used interchangeably to refer to the use of more than one therapy. The use of these terms does not restrict the order in which therapies are administered to an individual.

[0125] The compounds and pharmaceutical compositions described herein can be used in combination therapies having one or more additional therapeutic agents. For combination therapies having more than one active agent, where the active agent is in a single dosage form formulation, the active agent can be administered alone or in combination. Furthermore, an element can be administered before, simultaneously with, or after the administration of other agents.

[0126] When used in combination therapy with other agents, the "therapeutic effective amount" of the compounds and pharmaceutical compositions described herein, as well as one or more other agents, will depend on the type of drug used. The appropriate dosage of an approved drug is known and can be adjusted by someone skilled in the art based on the individual's condition, the type of condition being treated, and the amount of the compound described herein used. Where the dosage is not explicitly stated, an effective amount should be assumed.

[0127] In some embodiments, co-dosing or combination therapy encompasses the dosing of first and second amounts of a compound in a substantially simultaneous manner (such as in a single pharmaceutical composition, for example, capsules or tablets having first and second amounts in a fixed ratio, or multiple separate capsules or tablets). Additionally, such co-dosing also encompasses the sequential use of the compounds in any order.

[0128] When co-administration involves the separate administration of a first amount of Formula I compound and a second amount of additional therapeutic agent, the compounds are administered sufficiently close in time to achieve the desired therapeutic effect. For example, the time interval between the administrations that produce the desired therapeutic effect can range from minutes to hours, and this time interval can be determined taking into account the characteristics of each compound (such as potency, solubility, bioavailability, plasma half-life, and kinetic profile). For example, the Formula I compound and the second therapeutic agent can be administered in any order within approximately 24 hours, approximately 16 hours, approximately 8 hours, approximately 4 hours, approximately 1 hour, or approximately 30 minutes of each other.

[0129] Examples of other therapeutic agents that can be combined with, administered separately from, or in the same pharmaceutical composition as a compound of Formula I or a pharmaceutically acceptable salt thereof include, but are not limited to: (1) Endothelial-derived releasing factor (EDRF) or NO gas. (2) NO donors, including but not limited to nitrosothiols, nitrites, syl ketone imines, NONO salts (NONOate), N-nitrosamines, N-hydroxynitrosamines, nitrosimines, nitrotyrosine, diazacyclobutene dioxide, triazole 5-imine, oximes, hydroxylamines, N-hydroxyguanidines, hydroxyurea, or furazolidone. Some examples of these types of compounds include: trinitroglycerin esters (also known as GTN, nitroglycerin, nitroglycerin esters, and trinitroglycerin), nitrate esters of glycerol; sodium nitroprusside (SNP), in which the nitric oxide molecule is coordinated with iron metal to form a square bipyramidal complex; 3-(N-α-phosphoryl)sedone imine (SIN-1), an amphoteric compound formed by the combination of α-phosphoryl and sedone imine; S-nitroso-N-acetylopine (SNAP), an N-acetylated amino acid derivative with a nitrosothiol functional group; diethyltriamine / NO (DETA / NO), a nitric oxide compound covalently linked to diethyltriamine; and acetylsalicylic acid meta-nitrooxymethylphenyl ester. More specific examples of some of these NO donors include: typical nitro vasodilators, such as organic nitrates and nitrites, including nitroglycerin, amyl nitrite, isosorbide dinitrate, isosorbide 5-mononitrate, and nicorandil; isosorbide; 3-(N-α-linyl)stendone imine; lincidolamine hydrochloride ("SIN-1"); S-nitroso-N-acetylopine ("SNAP"); S-nitrosoglutathione (GSNO), sodium nitroprusside, S-nitrosoglutathione mono-ethyl-ester (GSNO-ester), 6-(2-hydroxy-1-methyl-nitrohydrazyl)-N-methyl-1-hexylamine, or diethylamine NONO salt. (3) Other substances that enhance cGMP concentration, including but not limited to protoporphyrin IX, eicosatetraenoic acid and phenylhydrazine derivatives. (4) Nitric oxide synthase acceptors, including but not limited to L-arginine, n-hydroxyguanidine analogs, such as N[G]-hydroxy-L-arginine (NOHA), 1-(3,4-dimethoxy-2-chlorobenzylmethylamino)-3-hydroxyguanidine and PR5 (1-(3,4-Dimethoxy-2-chlorobenzylmethylamino)-3-hydroxyguanidine); L-arginine derivatives (such as methylarg, methylNOHA, N-tert-butoxy- and N-(3-methyl-2-butenyl)oxy-L-arginine, canavalianic acid, ε-guanidine-hexanoic acid, guanidinebutamine, hydroxy-guanidinebutamine and L-tyramine aceto-L-arginine); N-alkyl-N'-hydroxyguanidine (such as N-cyclopropyl-N'-hydroxyguanidine and n-butyl-N'-hydroxyguanidine), N-aryl-N'-hydroxyguanidine (such as N-phenyl-N'-hydroxyguanidine and its para-substituted derivatives, which respectively carry -F, -Cl, -methyl, -OH substituents); guanidine derivatives, such as 3-(trifluoromethyl)propylguanidine. (5) Compounds that enhance eNOS transcription. (6) NO-independent heme-independent sGC activators, including but not limited to BAY 58-2667 (described in patent publication DE19943635); HMR-1766 (ataciguat, described in patent publication WO2000002851); S 3448 (2-(4-chloro-benzenesulfonamide)-4,5-dimethoxy-N-(4-(thio-lin-4-sulfonyl)-phenyl)-benzamide (described in patent publications DE19830430 and WO2000002851); and HMR-1069 (from Sanofi-Aventis). (7) Heme-dependent and NO-independent sGC stimulants, including but not limited to YC-1 (see patent publications EP667345 and DE19744026); riociguat (BAY 63-2521, Adempas®, described in DE19834044); nelociguat (BAY 60-4552, described in WO 2003095451); vericiguat (BAY 1021189, described in US8420656); BAY 41-2272 (described in DE19834047 and DE19942809); BAY 41-8543 (described in DE19834044); ectreciguat (described in WO 2003086407); CFM-1571 (Described in patent publication WO2000027394); A-344905, its acrylamide analog A-350619 and aminopyrimidine analog A-778935; [] Other sGC stimulants described in one of the following disclosures: US20090209556, US8455638, US20110118282 (WO2009032249), US20100292192, US20110201621, US7947664, US8053455 (WO2009094242), US20100216764, US8507512 (WO2010099054), US20110218202 (WO2010065275), US20130012511 (WO2011119518), US20130072492 (WO2011149921), US20130210798 (WO2012058132) and Tetrahedron Letters (2003), 44(48):8661-8663; and IW1973 (praliciguat), IW1701 (olinciguat) and CY6463 (previously IW-6463). (8) Compounds that inhibit the degradation of cGMP and / or cAMP, including but not limited to: PDE1 inhibitors; PDE2 inhibitors; PDE-3 inhibitors, such as amrinone, milrinone, enoximone, vesnarinone, pimobendan, and olprinone; PDE4 inhibitors, such as rolumilast; PDE5 inhibitors, such as sildenafil and related agents, such as avanafil, lodenafil, mirodenafil, and sildenafil citrate. citrate, tadalafil, vardenafil, and udenafil; alprostadil; dipyridamole and PF-00489791; PDE6 inhibitors, PDE9 inhibitors such as PF-04447943; PDE10 inhibitors such as PF-02545920 (PF-10); and PDE11 inhibitors. (9) Anticoagulants, including but not limited to: [Coumarins] (vitamin K antagonists), such as warfarin, cenocoumarol, phenprocoumon, and phenindione; Heparin and its derivatives, such as low molecular weight heparin, fondaparinux, and idraparinux; [Direct thrombin inhibitors], such as argatroban, lepirudin, bivalirudin, dabigatran, and ximelagatran; and Used to dissolve clots and break up arterial clots. [Tissue plasminogen activators], such as alteplase. (10) Antiplatelet drugs, including but not limited to topidogrel, ticlopidine, dipyridamole and aspirin. (11) Supplemental oxygen therapy. (12) α-1-adrenergic receptor antagonists, including but not limited to prazosin, indoramin, urapidil, bunazosin, terazosin and doxazosin; atrial natriuretic peptide (ANP), ethanol, histamine inducers, tetrahydrocannabinol (THC) and papaverine. (13) Bronchodilators, including but not limited to: [Short-acting] [β, 2, ] [Action activators], such as salbutamol or albuterol and terbutaline; [Long-lasting] [β, 2, ] [Accelerator] [(LABA)], such as salmeterol and formoterol; [Anticholinergic agents], such as pratropium and tiotropium; and Theophylline, bronchodilators, and phosphodiesterase inhibitors. (14) Corticosteroids, including but not limited to beclomethasone, methylprednisolone, betamethasone, prednisone, prednisolone, triamcinolone, dexamethasone, fluticasone, flunisolide, hydrocortisone; and corticosteroid analogs, such as budesonide. (15) Dietary supplements, including but not limited to omega-3 oils; folic acid, niacin, zinc, copper, ginseng root, ginkgo, pine bark, tribulus terrestris, arginine, oats, epimedium, Peruvian ginseng root, muira puama, saw palmetto and Swedish pollen; vitamin C, vitamin E, vitamin K2; testosterone supplements, transdermal testosterone patches; zoraxel, naltrexone, bremelanotide and melanotan II. (16) PGD2 receptor antagonists. (17) Immunosuppressants, including but not limited to cyclosporine, tacrolimus, rapamycin and other FK-506 immunosuppressants, mycophenolate, mycophenolate mofetil. (18) Nonsteroidal antiasthmatic agents, including but not limited to: [β2-] [Action agonists], such as terbutaline, isoproterenol, fenoterol, isoetharine, salbutamol, salmeterol, bitolterol, and pirbuterol; [β2-] [Accelerator] [-] [Corticosteroid combination], such as salmeterol-fluticasone, formoterol-budesonide, theophylline, cromoglycine (cromolyn), sodium cromoglycate, nedocromil, atropine, ipratropium, ipratropium bromide; and [Inhibitors of leukotriene biosynthesis], such as zileuton or veliflapon. (19) Nonsteroidal anti-inflammatory drugs (NSAIDs), including but not limited to: [Propionate derivatives], such as aminoprofen, benzoxaprofen, buprofen, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprazine, pirprofen, pranoprofen, sulprofen, thiamethoxam, and thioprofen; [Acetic acid derivatives], such as indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, thiophene, tolmetin, zidometacin, and zomepirac; [Fenamic acid derivatives], such as flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid, and tolfenamic acid; [Diphenylcarboxylic acid derivatives], such as diflunisal and flufenisal; [Xikang]( [oxicam], such as isoxicam, piroxicam, sudoxicam, and tenoxicam; [Salicylates], such as acetylsalicylic acid and sulfasalazine; and [Pyrazolinones], such as apazone, bezpiperylon, feprazone, mofebutazone, oxyphenbutazone, and phenylbutazone. (20) Cyclooxygenase-2 (COX-2) inhibitors, including but not limited to celecoxib, rofecoxib, valdecoxib, etoricoxib, parecoxib, and lumiracoxib; opioid analgesics, such as codeine, fentanyl, hydromorphone, levorphanol, meperidine, methadone, morphine, oxycodone, oxymorphone, propoxyphene, buprenorphine, butorphanol, dezocine, nalbuphine, and pentazocine; (21) Adrenergic neuron blocking agents, including but not limited to guanethidine and guanadrel. (22) Imidazolidine I-1 receptor agonists, including but not limited to rimenidine dihydrophosphate and mosonidin hydrochloride hydrate. (23) Potassium channel activators, including but not limited to pinacidil. (24) Dopamine D1 agonists, including but not limited to fenoldopam mesilate; other dopamine agonists, such as ibopamine, dopexamine and docarpamine. (25) 5-HT2 antagonists, including but not limited to ketanserin. (26) Vasopressin antagonists, including but not limited to tolvaptan. (27) Calcium channel sensitizers, including but not limited to levosimendan or activators such as nicorandil. (28) Adenylate cyclase activators, including but not limited to colforsin dapropate hydrochloride. (29) Positive myocardial systolic agents, including but not limited to digoxin and metildigoxin; metabolic cardiotonics, such as ubicarenone; and cerebral natriuretic peptides, such as nesiritide. (30) Medications used to treat erectile dysfunction, including but not limited to alprostadil, avitadil and phentolamine mesilate. (31) Medications used to treat Alzheimer's disease and dementia, including but not limited to: [Acetylcholinesterase inhibitors], such as galantamine, rivastigmine, donepezil, and tacrine; and [NMDA] [Receptor antagonists], such as memantine; and [Oxidoreductase inhibitors], such as idebenone. (32) Psychotropic drugs, including but not limited to: Ziprasidone, risperidone, olanzapine, valproate; [Dopamine] [D4] [Receptor antagonists], such as clozapine; [Dopamine] [D2] [Receptor antagonists], such as nemonapride; [Mixed Dopamine] [D1 / D2] [Receptor antagonists], such as zuclopenthixol; [GABA A] [Receptor modulators], such as carbamazepine; Sodium channel inhibitors, such as lamotrigine; Monoamine oxidase inhibitors, such as moclobemide and indeloxazine; and primavanserin and perospirone. (33) Medications used to treat movement disorders or symptoms, including but not limited to: [Catechols] [-O-] [Methyltransferase inhibitors], such as entacapone; Monoamine oxidase [B] [Inhibitors], such as selegiline; [Dopamine receptor modulators], such as levodopa; [Dopamine] [D3] [Receptor agonists], such as pramipexole; [Decarboxylase inhibitors], such as carbidopa; [Other dopamine receptor agonists], such as pergolide, ropinirole, and cabergoline; Rtigonide, istradefylline, talipexole; zonisamide and safinamide; and [Inhibitors of synaptic vesicle amine transporters], such as tetrabenazine. (34) [Medications used to treat mood or affective disorders or OCD, such as the following types:] [Tricyclic antidepressants], such as amitriptyline, desipramine, imipramine, amoxapine, nortriptyline, doxepin, and clomipramine; Selective serotonin reuptake inhibitors (SSRIs) [,]such as paroxetine, fluoxetine, sertraline, trazodone, and citralopram; [Atypical antidepressants], such as agomelatine; Selective norepinephrine reuptake inhibitor [(SNRI)], such as venlafaxine, reboxetine and atomoxetine; dopamine-induced antidepressants, such as bupropion and amineptine. (35) Drugs used to enhance synaptic plasticity, including but not limited to: [Nicotinic acid receptor antagonists], such as mecamylamine; and [mix] [5-HT] [Dopamine and norepinephrine receptor agonists], such as lurasidone. (36) Medications used to treat ADHD, such as amphetamine; 5-HT receptor modulators, such as vortioxetine; and α-2 adrenergic receptor agonists, such as chlorpromazine. (37) Nitric oxide synthase cofactors, including but not limited to tetrahydrobiopterin, dihydrobiopterin and sapropterin. (38) Blood glucose lowering agents (also known as blood glucose control agents or antidiabetic agents) include, but are not limited to: [Biguanide], such as metformin; [Sulfonylureas], such as glyburide, glybenclamide, glipizide, gliclazide, gliquidone, glimepiride, and combinations thereof with glimepiride, including atorvastatin calcium, meglinatide, tolbutamide, chlorpropamide, acesulfame potassium, and tolazimide; [α] [-] [Glucosidase inhibitors], such as acarbose, epalrestat, voglibose, and miglitol; [Insulin secretagogues], such as repaglinide, mitiglinide, and nateglinide; [Thiazolidinediones], such as rosiglitazone, troglitazone, ciglitazone, pioglitazone, empiglitazone, lobeglitazone sulfate, and balaglitazone; [DPP-4] [Inhibitor] [(] [or] [DPP-IV] [Inhibitor] [)], such as sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, alogliptin benzoate in combination with metformin or metformin hydrochloride, anagliptin, teneligliptin, atorvastatin calcium and glimepiride, empagliflozin in combination with linagliptin, gemigliptin, sitagliptin phosphate monohydrate in combination with pioglitazone hydrochloride, sitagliptin in combination with pioglitazone, sitagliptin in combination with atorvastatin calcium and (2 S,4) S)-1-[2-(1,1-dimethyl-3-sideoxy-3-pyrrolidin-1-yl-propylamino)acetyl]-4-fluoro-pyrrolidin-2-carboxynitrile (DBPR-108); [GLP-1] [Receptor] [Action promoters or incretin mimics], such as exenatide, dulaglutide, liraglutide, semaglutide, lixisenatide, lixisenatide in combination with insulin glargine, albiglutide, and pegapamodutide (TT-401), LY3298176 (a diglucose-dependent insulinotropic peptide (GIP) and GLP-1 receptor action promoter); [SGLT2] [Inhibitor] [(SGLT2i)], such as empagliflozin, empagliflozin in combination with linagliptin, empagliflozin in combination with metformin, ipragliflozin, ipragliflozin L-proline, tofogliflozin, sergagliflozin escargot, regoragliflozin escargot, ertugliflozin, ergagliflozin in combination with sitagliptin, ergagliflozin in combination with metformin, sotagliflozin, canagliflozin, canagliflozin in combination with metformin or metformin hydrochloride, dapagliflozin, dapagliflozin in combination with metformin or metformin hydrochloride and luseoglifozin, and dapagliflozin in combination with saxagliptin; [SGLT] [1] [Inhibitor or] [SGLT1] [and] [SGLT2] [Combinations of inhibitors], such as soggliflozin; [Insulin therapy] includes many types of insulin, such as insulin lispro, insulin degludec, insulin lispro, insulin aspart, insulin glargine, insulin detemir, protamine insulin, mixed insulin (human insulin containing both rapid-acting (soluble) and long-acting (protamine) insulin), insulin degludec in combination with insulin aspart, human (rDNA-derived) inhaled powder insulin, recombinant human insulin, liver-guided vesicular insulin, insulin tregopi (IN-105), insulin degludec in combination with liraglutide, insulin peglispro (LY-2605541), and nodlin; and [Tolimedone] [tolimidone] (lyn kinase activator). (39) Antihypertensive drugs (also known as antihypertensive agents), including but not limited to: [Diuretics], such as thiazide diuretics, chlorothiazide, chlorothiazide, hydrochlorothiazide, benzylfluorothiazide, cyclopenthiazine, methazothiazide, polythiazides, quinetrazolone, xipamide, metolazone, indapamide, cicletanine, furosemide, toresamide, amiloride, spironolactone, potassium canileate, eplerenone, triamterene, acetazolamide, and carperitone; [β] [Blockers], such as acebutolol, atenolol, metoprolol, and nebivolol; [Angiotensin-converting enzyme] [(ACE)] [Inhibitors], such as thiol-containing agents (e.g., captopril, zofenopril), dicarboxylate-containing agents (e.g., enalapril, quinapril, ramipril, perindopril, lisinopril, and benazepril), phosphonate-containing agents (e.g., fosinopril), and naturally occurring ACE inhibitors. Agents (e.g., tyrosinase, lactokine, lactotripeptide Val-Pro-Pro and Ile-Pro-Pro), alacepril, delapril, cilazapril, imidapril, temocapril, moexipril, lisinopril, combinations of lisinopril and hydrochlorothiazide, trandolapril, and spirapril); [Angiotensin] [II] [Receptor blockers] [(ARB)] [,] Such as candesartan, losartan, losartan potassium-hydrochlorothiazide, valsartan, candesartan cilexetil, eprosartan, irbesartan, telmisartan, olmesartan medull (or olmesartan), azilsartan medoxomil, azilsartan, and amlodipine besylate in combination with irbesartan. besylate, azilsartan in combination with amlodipine besylate, cilnidipine in combination with valsartan, fimasartan, irbesartan in combination with atorvastatin, irbesartan in combination with chlorothiazide, losartan potassium in combination with hydrochlorothiazide and / or amlodipine besylate, pratosartan, atorvastatin calcium in combination with losartan potassium, nifedipine and candesartan medoxomil, sacubitril in combination with valsartan or LCZ-696, angiotensin AT2 antagonists and TAK-591, and olmesartan medumid; Endothelin receptor antagonists [(ERA)] [,] Such as atrasentan, bosentan, sitaxentan, ambrisentan, actelion-1 (macitentan), D-trp-D-asp-L-pro-D-Val-L-leu (BQ-123), sparsentan, and tezosentan disodium. [Mineralocorticoid receptor antagonists] [(MRA)] [,]such as spironolactone, spironolactone combined with ampicillin hydrochloride, apararenone or MT-3995, eplerenone, and finerenone (BAY-94-8862); [Calcium channel blockers], such as amlodipine, aranidipine, azelnidipine, barnidipine, benidipine, cilnidipine, clevidipine, diltiazem, efonidipine, felodipine, lacidipine, lercanidipine, and manidipine. e) Nicardipine, Nifedipine, Nivadipine, Nimodipine, Nisoldipine, Nitrondidipine, Pranidipine, Isradipine, Verapamil, Gallopamil, Diltiazem, Mibefradil, Bepridil, Fluspirilene, and Fendiline; [Renin inhibitors], such as aliskiren; [α] [Blockers], such as doxazosin and prazosin; [α] [-] [β] [Blockers], such as carvedilol and labetalol; [Central nervous system drugs], such as clonidine, guanfacine, and methyldopa; [Vasodilators], such as nitroglycerin, hydralazine, and minoxidil; and [Aldosterone antagonists], such as fenestrone, spironolactone and eplerenone. (40) Antilipidemia agents, including but not limited to: [Startrine] [(s)] [tatins] Such as atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin; [Combinations of statins with another medication], such as amlodipine / atorvastatin, aspirin / pravastatin, ezetimibe / simvastatin, niacin / simvastatin, lovastatin / niacin, simvastatin / sitagliptin, and atorvastatin / ezetimibe; [Cellulose esters or cellulose acid derivatives]. Examples include, but are not limited to, fenofibrate, gemfibrozil, bezafibrate, ciprofibrate, clinofibrate, and clofibrate; [Nicotinic acid] (or nicotinic acid); [Bile acid sequestrants], such as cholestyramine, colesevelam, colestilan, and colestipol; [Ezemab, Lometab] [(l)] [omitapide] [、] [Phytosterols or Orlistat] [(orlistat)] [;] [and] [PCSK9] [Inhibitors], such as alicurumab and evolocumab; (41) Endopeptidase inhibitors (also known as endopeptidase inhibitors or NEP inhibitors or neprilysin inhibitors), including but not limited to sacubitril, or a combination of sacubitril and valsartan; neprilysin inhibitors, such as TD-1439 or TD-0714 under development. (42) Kidney-protective drugs, including but not limited to: Bardoxolone [ACE] [Inhibitors], such as captopril; [ARB] [,]such as losartan or ibesartan; [SGLT2] [Inhibitors], such as canagliflozin, [GLP1] [Receptor agonists] [;] [MRA] [,]such as fenestrone; [ERA] [,]such as atrazotan; and Inhibitors of apoptosis signal-regulated kinase 1 (ASK1), such as selonsertib. (43) Hydroxyurea (HU, hydroxyurea). (44) Anti-sickle cell formation agents, including but not limited to hydroxyurea, voxelotor or GBT-440. (45) Anti-adhesion therapy, including but not limited to blocking antibody adhesion to P-selectin, E-selectin, VLA-4, and VCAM-1. (46) Glutamic acid. (47) Erythropoietin (EPO), also known as hematopoietin, includes all its forms, such as exogenous erythropoietin, recombinant human erythropoietin (rhEPO) or other erythropoiesis stimulants (ESAs), two examples being epoetin alfa and epoetin beta. (48) Antibiotics, including but not limited to: [Penicillin and its derivatives], including but not limited to penicillin, amoxicillin, ampicillin, azlocillin, cloxacillin, penicillin G, penicillin V, procaine penicillin or benzathine penicillin, etc. Cephalosporins, such as cephalexin, cefadroxil, cefaclor, cefuroxime, and cefixime; [Macrocyclic lactones], such as erythromycin, clarithromycin, azithromycin, and roxithromycin; [Tetracyclines and their derivatives], such as demeclocycline, doxycycline, minocycline, oxytetracycline, and tetracycline; [Sulfamethamides] include, but are not limited to, mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfamethoxazole, sulfamethoxazole, sulfasalazine, trimethoprim-sulfamethoxazole (co-trimoxazole), and sulfamethoxazole; and Quinolinones, including but not limited to ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, ofloxacin, and nalidixic acid. (49) FXR agonists, including but not limited to obeticholic acid, cenicriviroc, emricasan, GR-MD-02, slonseter and elafibranor. (50) Thyroid receptor-β agonists, including but not limited to MGL-3196. (51) Acetyl-CoA carboxylase inhibitors, including but not limited to GS-0976. (52) Treatments for mitochondrial disorders, including but not limited to vitamins and supplements, including coenzyme Q10; B complex vitamins, especially thiamine (B1) and riboflavin (B2); alpha fatty acids; L-carnitine; creatine; citrulline and L-arginine. (53) Therapeutic agents for epilepsy or seizures, including but not limited to phenytoin, valproic acid, phenobarbital, lamotrigine, carbamazepine, topiramate, oxcarbazepine, zonisamide, gabapentin, levetiracetam, pregabalin, clonazepam, lacosamide, rufinamide, and vigabatrin.

[0130] [, , ] [, Packages and assemblies , ] [, , ] Depending on the method of drug administration, pharmaceutical compositions (or formulations) for use may be packaged in various ways. Generally, articles for dispensing include containers in which pharmaceutical formulations are stored in a suitable form. Suitable containers are well known to those skilled in the art and include materials such as bottles (plastic and glass), capsules, ampoules, plastic bags, metal canisters, and the like. Containers may also include tamper-evident fittings to prevent easy access to the contents. Additionally, the container is labeled with a description of its contents. The label may also include appropriate warnings.

[0131] The compounds and pharmaceutical formulations described herein may be contained in a kit. A kit may include two or more pharmaceutical agents, individually packaged or prepared in single or multiple doses, or in combination of two or more pharmaceutical agents, packaged or prepared in single or multiple doses. Thus, one or more pharmaceutical agents may be present in a first container, and the kit may include one or more pharmaceutical agents in a second container, as appropriate. One or more containers are placed within a package, and the package may include, as appropriate, instructions for dosing or dosing. The kit may include additional components, such as syringes or other components for dosing the pharmaceutical agents and diluents, or other components for preparation. Therefore, the kit may contain: a) a pharmaceutical composition comprising the compounds described herein and a pharmaceutically acceptable carrier, mediator, or diluent; and b) a container or package. The kit may include, as appropriate, instructions for use of the pharmaceutical composition in one or more of the methods described herein (e.g., for the prevention or treatment of one or more of the diseases and conditions described herein). The kit may, where appropriate, include a second pharmaceutical composition comprising one or more additional pharmaceutical agents, pharmaceutically acceptable carriers, mediators, or diluents as described herein for co-therapeutic use. The pharmaceutical composition comprising the compounds described herein and the second pharmaceutical composition contained in the kit may, where appropriate, be combined within the same pharmaceutical composition.

[0132] [] [Example] [] All references provided in the examples are incorporated herein by reference. As used herein, all abbreviations, symbols, and rules are consistent with their use in contemporary scientific literature. See, for example, Janet S. Dodd, ed., The ACS Style Guide: A Manual for Authors and Editors, 2nd ed., Washington, DC: American Chemical Society, 1997, which is incorporated herein by reference in its entirety.

[0133] Various embodiments of the present invention are described below.

[0134] The definitions of the abbreviations used in the examples section are provided in the table below. abbreviation Word groups or phrases app. obvious Conc. concentration DMF N,N-Dimethylformamide DMSO Dimethicone DMSO-d6 Deuterated dimethyl sulfide dppf 1,1'-bis(diphenylphosphine)ferrocene dba Dibenzylideneacetone ES+ Positron spray ionization EtOAc Ethyl acetate equiv. equivalent FA Formic acid h Hour HEK Human embryonic kidney HPLC High performance liquid chromatography i-PrOH Isopropanol LC-MS Liquid chromatography-mass spectrometry MeOH methanol mL milliliters MHz million Hertz MS Mass spectrometry NMR Nuclear magnetic resonance No. Compound numbering Ph Phenyl ppm parts per million RP-HPLC Reverse-phase high performance liquid chromatography TFA Trifluoroacetic acid THF Tetrahydrofuran

[0135] [Synthesis Section] [, , ] [Example] [1] [:] [Composite] [I] [Compound] [, , ] This invention also provides a method for synthesizing compounds of formula I, which represents another embodiment of the invention. Compounds of formula I of the invention can be prepared according to the general and specific syntheses described herein, synthetic procedures reported in the chemical literature, or methods known to those generally skilled in the art. As will be understood by those generally skilled in the art, the optimal reaction conditions that can be determined during experiments can be varied based on the type of reaction and the specific reagents used in the reaction. Therefore, unless specifically described, reaction conditions such as pressure, temperature, relative ratios of reagents, solvents, and reaction time can be easily selected and modified by those generally skilled in the art without any improper experimentation. The compounds and intermediates of this invention can be prepared by purification methods known to those skilled in the art. These methods include, but are not limited to, silica gel chromatography, recrystallization, reversed-phase HPLC (RP-HPLC), and supercritical fluid chromatography (SFC). RP-HPLC purification can be achieved using a suitable gradient selected from 0% to 100% acetonitrile / water containing an additive such as 0.1% TFA or FA on a suitable reversed-phase column (e.g., Waters XBridge OBD C18, 5 µm, 19 × 150 mm). Non-mirror image isomers can be separated by silica gel chromatography, RP-HPLC, or palmar HPLC. Discrete mirror image isomers can be obtained by resolving mixtures of mirror image isomers using palmar HPLC. The reaction progress can be monitored by methods known to those skilled in the art, such as thin-layer chromatography, reversed-phase HPLC, or tandem reversed-phase HPLC-mass spectrometry (LC-MS).

[0136] The starting materials used in the synthesis described herein were purchased from commercial sources or can be prepared by a person generally familiar with this technique using methods reported in chemical literature or mentioned herein.

[0137] The general methods described herein can be used to prepare compounds of formula I. The general and specific methods described herein are provided as illustrations for carrying out the invention. Therefore, they are not intended to impose any limitation on the subject matter and scope of the claimed compounds of the invention.

[0138] All references provided in the examples are incorporated herein by reference. As used herein, all abbreviations, symbols, and rules are consistent with their use in contemporary scientific literature. See, for example, GM Banik, G. Baysinger, PV Kamat, NJ Pienta, eds., The ACS Guide to Scholarly Communication, Washington, DC: American Chemical Society, 2020 (https: / / pub.acs.org / doi / book / 10.1021 / acsguide), which is incorporated herein by reference in its entirety.

[0139] [Example] [1] [Compound Synthesis] The compounds disclosed herein can be prepared from corresponding nitrile intermediates, for example, using the general procedure (General Procedure C) described below:

[0140] [General Program] [C] The compounds of the present invention can be prepared by means of their corresponding nitriles via procedures similar to those described herein. Nitriles with different substitution modes can be prepared by means of the procedures described in WO2015187470, WO2016081668, WO2017197555, WO2017200825, WO2018 / 045276A1 and WO2019 / 126354A1.

[0141] The following nitrile intermediates were prepared according to the procedures described in WO2018 / 045276A1 and WO2019 / 126354A1. Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. 8-Benzylimidazo[1,2-a]pyridine-6-carboxynitrile; 8-(3-fluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(2-fluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(2,3-difluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(2,5-Difluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(3-fluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(3,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(3,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile; 8-(2,5-difluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-carboxynitrile; 8-(3-fluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-carboxynitrile; 8-(3,5-difluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-carboxylonitrile; 8-(2,3-difluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-carboxylonitrile; 8-(2,5-difluoro-4-methylbenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-carboxylonitrile.

[0142] The procedures for synthesizing compounds I-1 to I-20 are described below. The procedures for synthesizing compounds I-20 to I-26 are described in patent application publication WO2019 / 126354.

[0143] [] [General Program] [A] [,] [Suitable for synthesizing compounds] [I-1] [] The title compound was synthesized in two steps:

[0144] [Steps] [1] [:] Synthesis of 8-(3-fluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate Add 0.50 N sodium methoxide in methanol (0.17 mL, 0.087 mmol, 0.10 equivalent) to a solution of 220 mg, 0.87 mmol, 1.0 equivalent) in methanol (5.0 mL) (Note: stoichiometric amounts or excess sodium methoxide may also be used). After stirring at ambient temperature for 6 h, add ammonium chloride (280 mg, 5.2 mmol, 6.0 equivalent) and stir the reaction mixture for 16 h. Concentrate the reaction mixture under vacuum, dilute with 20 mL of half-saturated NaHCO3 solution and extract with 2 × 20 mL of CH2Cl2 / iPrOH (5:1). The combined organic phases were dried with sodium sulfate, filtered, and concentrated to obtain a crude formamidine product, which was a light brown foamy solid. It was used in the next step without further purification. LC / MS ES + m / z = 270.2 [M+H]+.

[0145] [Steps] [2]: Synthesis of 5-fluoro-2-(8-(3-fluorobenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol Add (Z)-3-ethoxy-2-fluoro-3-sideoxyprop-1-en-1-ol sodium salt (490 mg, 3.1 mmol, 4.0 equivalent) to a suspension of 8-(3-fluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate (210 mg, 0.79 mmol, 1.0 equivalent) in ethanol (7.0 mL). Heat the reaction mixture in a sealed vial at 90 °C for 2.5 h. After cooling to ambient temperature, add 1.0 N HCl aqueous solution (3.1 mL, 3.1 mmol, 4.0 equivalent). The resulting mixture was concentrated under vacuum, diluted with water (50 mL), adjusted to pH 6 with saturated NaHCO3 solution, and extracted with 2 × 50 mL CH2Cl2 / iPrOH (5:1). The combined organic phase was dried over sodium sulfate, filtered, and concentrated. The crude material was purified by silica gel chromatography (0 to 15% acetonitrile / methanol (7:1) in CH2Cl2) to give the title compound as a light brown solid (180 mg, 64% yield, in 2 steps). ¹H NMR (500 MHz, DMSO-d⁶) δ (ppm) 13.1–12.5 (single pair, ¹H, tautomer), 9.46 (s, ¹H), 8.30 (s, ¹H), 8.26–8.00 (single pair, ¹H, tautomer), 7.90 (s, ¹H), 7.50 (m, ¹H), 7.41 (m, ¹H), 7.32 (m, ¹H), 7.02 (app. t, ¹H), 4.53 (s, ²H).

[0146] [Compound] [I-2] 2-(8-benzylimidazo[1,2-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol was synthesized as a white solid according to general procedure A. [(] [Compound] [I-] [2)](25 mg, 14% total yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, methanol-d⁴) δ (ppm) 9.30 (s, ¹H), 8.09 (s, ¹H), 7.99 (d, ¹H), 7.80 (s, ¹H), 7.40 (d, ²H), 7.17 (t, ²H), 7.07 - 7.11 (m, ¹H), 4.52 (s, ²H).

[0147] [Compound] [I-4] 2-(8-(2,3-difluorobenzyl)imidazo[1,2-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol (compound 4) was synthesized as a light brown solid according to general procedure A (150 mg, 67% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods were varied as needed. ¹H NMR (500 MHz, acetone-d6) δ (ppm) 10.6 (s, 1 H), 9.34 (s, 1 H), 8.16 (s, 1 H), 7.92 (s, 1 H), 7.78 (s, 1 H), 7.20 (t, 1 H), 7.09 (q, 1 H), 7.00 (q, 1 H), 4.60 (s, 2 H).

[0148] [] [Compound] [I-6] 5-fluoro-2-(8-(3-fluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol, a pale yellow solid, was synthesized according to general procedure A. [Compound] [I-6] (200 mg, 54% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d₆) δ (ppm) 12.9 (br. s, ¹H), 9.44 (s, ¹H), 8.29 (s, ¹H), 8.16 (br. s, ¹H), 7.89 (s, ¹H), 7.39 (d, ¹H), 7.27 (d, ¹H), 7.17 (t, ¹H), 4.48 (s, ²H), 2.14 (s, ³H).

[0149] [] [Compound] [I-7] 2-(8-(3,5-difluorophenylmethyl)imidazo[1,2-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol, a yellow solid, was synthesized according to general procedure A. [Compound] [I-7]) (190 mg, 57% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d₆) δ (ppm) 13.0 (br. s, ¹H), 9.47 (s, ¹H), 8.31 (s, ¹H), 8.22 (br. s, ¹H), 7.91 (s, ¹H), 7.36 (br. s, ²H), 7.07 (t, ¹H), 4.53 (s, ²H).

[0150] [Compound] [I-3] 5-fluoro-2-(8-(3,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyrazin-6-yl)pyrimidin-4-ol, a white solid, was synthesized according to general procedure A. [Compound] [I-3]) 67 mg, 34% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, chloroform-d) δ (ppm) 11.1 (br. s, ¹H), 9.14 (s, ¹H), 8.00–7.91 (m, ²H), 7.87 (s, ¹H), 7.00 (d, ²H), 4.56 (s, ²H), 2.14 (s, ³H).

[0151] [Compound] [I-14] The title compound was synthesized in two steps:

[0152] [step] [1] [:] Synthesis of 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carbamate A solution of 0.50 N sodium methoxide in methanol (18 mL, 8.9 mmol, 1.0 equivalent) was added to a suspension of 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile (2.5 g, 8.9 mmol, 1.0 equivalent) in methanol (44 mL). After stirring at ambient temperature for 4 h, a further portion of methanol containing 0.50 N sodium methoxide (5.3 mL, 2.7 mmol, 0.3 equivalent) was added, and stirring continued for another 2 h. Ammonium chloride (470 mg, 8.9 mmol, 1.0 equivalent) was then added. After 16 h, the reaction mixture was concentrated under vacuum, suspended in a saturated aqueous solution of NaHCO3, and stirred for 20 min. The solid was collected by filtration and washed with 3 volumes of water and 2 volumes of ether. The crude product was resuspended in 100 mL of acetonitrile under heating, diluted with ether, and filtered. The filter cake was washed with 3 volumes of ether and dried to give a light brown solid (2.2 g, 83% yield). It was used in the next step without further purification. LC / MS ES + m / z = 302.1 [M+H]+.

[0153] [step] [2] [:] Synthesis of 5-fluoro-2-(8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol Add (Z)-3-ethoxy-2-fluoro-3-sideoxyprop-1-en-1-ol sodium chloride (2.9 g, 19 mmol, 3.0 equivalent) to ethanol (31 mL) containing 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carbamate (1.9 g, 6.2 mmol, 1.0 equivalent). Heat the solution in a sealed container at 90 °C for 18 h. After cooling to ambient temperature, add 7.4 mL of 2.5 N ethanol-HCl solution (19 mmol, 3.0 equivalent). Concentrate the resulting mixture under vacuum and suspend it in acetonitrile (100 mL) while heating. After slight cooling, add ether (100 mL) and stir the mixture for 10 min. The solid was collected by filtration and washed with 3 volumes of ether. The resulting solid was then suspended in water, stirred for 1 h, and filtered. The crude material was purified by preparative reversed-phase HPLC (10% to 70% acetonitrile / water with 0.1% trifluoroacetic acid as an additive). The impure solvent was further purified by preparative reversed-phase HPLC (10% to 50% acetonitrile / water with 0.1% trifluoroacetic acid as an additive) to give the title compound (840 mg, 37% yield) as a grayish-white solid. 1H NMR (500 MHz, methanol-d 4) δ (ppm) 9.43 (s, 1 H), 8.21 (s, 1 H), 8.08 (br. s, 1 H), 7.89 (s, 1 H), 7.09 (m, 1 H), 7.00 (m, 1 H), 4.63 (s, 2 H), 2.23 (s, 3 H).

[0154] [Compound] [I-14] [Of] [Na+] [salt] [] Under a nitrogen atmosphere, 5-fluoro-2-(8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol ( [Compound] [I-14] A solution of 0.50 N sodium methoxide in methanol (54 mL, 27 mmol) was added to a grayish-white suspension of [,] 10 g (27 mmol) in 450 mL of anhydrous MeOH. After simple sonication, the resulting pale yellow solution was stirred at ambient temperature for 15 min and concentrated to dryness under vacuum. The solid was resuspended in 250 mL of ether by sonication and concentrated (twice). The resulting solid was resuspended in 650 mL of ether and stirred at ambient temperature for 3 h. The solid was collected by vacuum filtration and washed with ether (3 × 100 mL). After drying on the filter overnight, the product salt was dried in a vacuum oven at 45 °C for 4 days to give sodium 5-fluoro-2-(8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-olide (11 g, 99% yield) as a white solid. 1H NMR (500 MHz, D 2O) (ppm) 8.92 (s, 1 H), 7.99 (d, 1 H), 7.97 (d, 1 H), 7.70 (d, 1 H), 6.98 (dd, 1 H), 6.86 (dd, 1 H), 4.48 (s, 2 H), 2.14 (s, 3 H).

[0155] [] [Compound] [I-11] According to general procedure A, 5-fluoro-2-(8-(3-fluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-yl)pyrimidin-4-ol, a golden solid, was synthesized. [Compound] [I-11])(61 mg, 23% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d6) δ (ppm) 13.3 (br. s, ¹H), 9.60 (s, ¹H), 8.86 (s, ¹H), 8.24–8.27 (m, ¹H), 7.32–7.47 (m, ³H), 7.03–7.06 (m, ¹H), 4.59 (s, ²H).

[0156] [] [Compound] [I-13] 2-(8-(3,5-difluorobenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol, a brown solid, was synthesized according to general procedure A. [Compound] [I-13]) (57 mg, 17% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d 6) δ (ppm) 13.2 (br. s, ¹H), 9.61 (s, ¹H), 8.87 (s, ¹H), 8.25 (s, ¹H), 7.33 (d, ²H), 7.10 (t, ¹H), 4.60 (s, ²H).

[0157] [] [Compound] [I-10] 2-(8-(2,3-difluorophenylmethyl)-[1,2,4]triazolo[1,5-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol, a pale yellow solid, was synthesized according to general procedure A. [Compound] [I-10] (85 mg, 16% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d6) δ (ppm) 13.0 (br. s, ¹H), 9.62 (s, ¹H), 8.85 (s, ¹H), 8.23 ​​(s, ¹H), 7.29–7.37 (m, ²H), 7.09–7.16 (m, ¹H), 4.68 (s, ²H).

[0158] [] [Compound] [I-12] 2-(8-(2,5-difluorophenylmethyl)-[1,2,4]triazolo[1,5-a]pyridine-6-yl)-5-fluoropyrimidin-4-ol, a grayish-white solid, was synthesized according to general procedure A. [Compound] [I-12]) (75 mg, 57% yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, methanol-d⁴) δ (ppm) 9.67 (s, ¹H), 8.69 (s, ¹H), 8.12 (d, ¹H), 7.25 (m, ¹H), 7.13 (m, ¹H), 7.02 (m, ¹H), 4.73 (s, ²H).

[0159] [Compound] [I-19] 5-fluoro-2-(8-(2,5-difluoro-4-methylbenzyl)-[1,2,4]triazolo[1,5-a]pyridine-6-yl)pyrimidin-4-ol, a light brown solid, was synthesized according to general procedure A. [Compound] [I-19] (140 mg, 66% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d6) δ (ppm) 13.1 (br. s, ¹H), 9.61 (s, ¹H), 8.83 (s, ¹H), 8.26 (br. s, ¹H), 7.35 (br. s, ¹H), 7.17 (m, ¹H), 4.57 (s, ²H), 2.18 (s, ³H).

[0160] [] [General Program] [B] [,] [Suitable for synthesizing compounds] [I-16] [] The title compound was synthesized in two steps:

[0161] [Steps] [1]: Synthesis of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate Add a solution of 0.50 N sodium methoxide in methanol (3.6 mL, 1.8 mmol, 1.0 equivalent) to a suspension of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile (490 mg, 1.8 mmol, 1.0 equivalent) in methanol (5.0 mL) (Note: catalytic amounts or excess sodium methoxide may also be used). After stirring at ambient temperature for 3 h 45 min, add ammonium chloride (970 mg, 18 mmol, 10 equivalent) and stir the reaction mixture for 20 h. Concentrate the resulting mixture under vacuum to approximately 2 mL and dilute with EtOAc (20 mL) and 10% NaHCO3 aqueous solution (10 mL). After stirring for 15 min, the product was collected by filtration, washed with water (10 mL), and dried under vacuum to give the title compound (420 mg, 80% yield) as a grayish-white solid. LC / MS ES+m / z = 287.9 ​​[M+H]+.

[0162] [Steps] [2] [:] Synthesis of 5-chloro-2-(8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol A solution of 0.50 N sodium methoxide in methanol (1.4 mL, 0.70 mmol) was added to a suspension of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate (100 mg, 0.35 mmol) and ethyl 2-chloro-3-sideoxypropionate (110 mg, 0.70 mmol) in methanol (1.7 mL). The reaction mixture was heated in a sealed vial at 65 °C for 2.5 h. After cooling to ambient temperature, the resulting mixture was concentrated under vacuum, diluted with water (10 mL), adjusted to pH 3 with 6.0 N HCl aqueous solution, and extracted with 2 × 15 mL CH₂Cl₂ / iPrOH (8:1). The combined organic phase was dried with sodium sulfate, filtered, and concentrated. The crude material was purified by silica gel chromatography (0% to 20% acetonitrile / methanol (7:1) in CH2Cl2) and then further purified by silica gel chromatography (20% to 100% EtOAc / CH2Cl2) to give the title compound (37 mg, 28% yield) as a grayish-white solid. 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 12.6 (br. s, 1 H), 9.54 (s, 1 H), 8.36 (br. s, 1 H), 8.32 (s, 1 H), 7.89 (s, 1 H), 7.45 (br. s, 1 H), 7.25 (m, 1 H), 7.12 (m, 1 H), 4.58 (s, 2 H).

[0163] [] [Compound] [I-17] 5-Chloro-2-(8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol, a light brown solid, was synthesized according to general procedure B. [Compound] [I-17] (5.2 mg, 2.2% gross yield). Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods may be modified as needed. ¹H NMR (500 MHz, DMSO-d₆) δ (ppm) 9.47 (s, ¹H), 8.45 (s, ¹H), 8.22 (d, ¹H), 7.94 (s, ¹H), 7.23 (dd, ¹H), 7.16 (dd, ¹H), 6.72 (d, ¹H), 4.56 (s, ²H), 2.17 (br s, ³H). LC / MS ES⁻¹ + m / z = 388.0 [M⁺H]⁺.

[0164] [] [Compound] [I-15] The title compound was synthesized in two steps:

[0165] [step] [1]: Synthesis of 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carbamate 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carbamate (840 mg, 76% yield) was synthesized as a light brown solid according to step 1 of general procedure A or B. Reaction conditions (such as reagent ratios, temperature, and reaction time) and purification methods were modified as needed. LC / MS ES + m / z = 302.0 [M+H]+.

[0166] [step] [2]: Synthesis of 2-(8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol Huenig's base (0.63 mL, 3.6 mmol) was added to a suspension of 8-(2,5-difluoro-4-methylbenzyl)imidazo[1,2-a]pyridine-6-carbamate (360 mg, 1.2 mmol) and methyl 3-methoxyacrylate (0.39 mL, 3.6 mmol) in ethanol (6.0 mL). The reaction mixture was heated in a sealed vial at 90 °C for 3 h. After cooling to ambient temperature, the mixture was treated with 2.5 N ethanol-HCl solution (1.4 mL, 3.6 mmol) and concentrated to dryness. The crude material was purified by silica gel chromatography (0% to 20% acetonitrile / methanol (7:1) in CH2Cl2) and then purified again by silica gel chromatography (0 to 15% MeOH / CH2Cl2) to give the title compound (120 mg, 28% yield) as a light brown solid. 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 11.9 (br. s, 1 H), 9.52 (s, 1 H), 8.31 (s, 1 H), 8.07 (br. d, 1 H), 7.89 (s, 1 H), 7.37 (dd, 1 H), 7.16 (dd, 1 H), 6.38 (br. d, 1 H), 4.54 (s, 2 H), 2.18 (s, 3 H).

[0167] [] [Compound] [I-8] The title compound was synthesized in two steps:

[0168] [Steps] [1]: Synthesis of 8-(2-fluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate 8-(2-fluorobenzyl)imidazo[1,2-a]pyridine-6-carboxamide (5.1 g, 91% yield) was synthesized as a creamy yellow solid according to step 1 of general procedure A or B. Reaction conditions (e.g., reagent ratios, temperature, and reaction time) and purification methods were modified as needed. LC / MS ES + m / z = 270.2 [M+H]+.

[0169] [Steps] [2]: Synthesis of 2-(8-(2-fluorobenzyl)imidazo[1,2-a]pyridine-6-yl)-5-methylpyrimidin-4-ol Potassium bicarbonate (220 mg, 2.2 mmol) was added to a solution of 8-(2-fluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate (400 mg, 1.5 mmol) and ethyl 2-methyl-3-sideoxypropionate (230 mg, 1.8 mmol) in t-BuOH (9.9 mL). The reaction mixture was heated to reflux for 2 h. After cooling to ambient temperature, water was added, and the product was collected by filtration and dried to give the title compound (410 mg, 82% yield) as a creamy yellow solid. 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 11.6 (br. s, 1 H), 9.48 (s, 1 H), 8.30 (s, 1 H), 7.94 (br s, 1 H), 7.88 (s, 1 H), 7.48 (app. t, 1 H), 7.29 (m, 1 H), 7.19 (m, 1 H), 7.11 (app. t, 1 H), 4.60 (s, 2 H), 1.98 (s, 3 H).

[0170] [Compound] [I-9] 9.0 mL of acetonitrile-THF (2:1) containing 5-fluoro-2-(8-(2-fluorobenzyl)imidazo[1,2-a]pyridyl-6-yl)pyrimidin-4-ol (230 mg, 0.67 mmol) was treated with sodium bicarbonate (84 mg, 1.0 mmol) and Selectfluor™ (350 mg, 1.0 mmol) and heated at 50 °C. Additional sodium bicarbonate (42 + 28 mg) and Selectfluor™ (180 + 120 mg) were added during the experiment. After a total of 49 h, the reaction mixture was cooled to ambient temperature and 20 mL of water was added. The resulting mixture was acidified to pH 3 with 1.0 N HCl aqueous solution and extracted with 2 × 25 mL of EtOAc. The combined organic phase was dried with sodium sulfate, filtered, and concentrated. The crude material was purified by silica gel chromatography (0% to 20% acetonitrile / methanol (7:1) in CH2Cl2) and further purified by preparative reversed-phase HPLC (15% to 65% acetonitrile / water with 0.1% formic acid as an additive) to give the title compound (23 mg, 9.7% yield) as a light brown solid. 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 12.6 (br. s, 1 H), 8.98 (s, 1 H), 8.19 (br. s, 1 H), 7.74 (d, 1 H), 7.48 (app. t, 1 H), 7.28 (m, 1 H), 7.19 (m, 1 H), 7.10 (app. t, 1 H), 4.56 (s, 2 H).

[0171] [] [Compound] [I-5] 10 mL of acetonitrile-THF (1:1) containing 5-fluoro-2-(8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol (200 mg, 0.56 mmol) was treated with sodium bicarbonate (94 mg, 1.1 mmol) and Selectfluor™ (400 mg, 1.1 mmol) and heated at 50 °C. Additional sodium bicarbonate (3 × 47 mg) and Selectfluor™ (3 × 200 mg) were added during the experiment. After a total of 74 h, the reaction mixture was cooled to ambient temperature and 40 mL of water was added. The resulting mixture was acidified to pH 3 with 1.0 N HCl aqueous solution and extracted with 2 × 40 mL of CH₂Cl₂ / iPrOH (6:1). The combined organic phase was dried with sodium sulfate, filtered, and concentrated. The crude material was purified by silica gel chromatography (0 to 20% acetonitrile / methanol (7:1) in CH2Cl2), preparative reversed-phase HPLC (10% to 70% acetonitrile / water with 0.1% TFA as an additive), and final column chromatography (20% to 100% EtOAc / hexane) to give the title compound as a white solid (24 mg, 11% yield). 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 12.8 (br. s, 1 H), 8.99 (s, 1 H), 8.20 (br. s, 1 H), 7.75 (d, 1 H), 7.42 (m, 1 H), 7.25 (m, 1 H), 7.13 (m, 1 H), 4.54 (s, 2 H).

[0172] [Compound] [I-18] The title compound was synthesized in 5 steps:

[0173] [Steps] [1]: Synthesis of 6,8-dibromo-3-fluoroimidazole[1,2-a]pyridine [] 40 mL of acetonitrile containing 2.4 g (8.7 mmol) of 6,8-dibromoimidazolo[1,2-a]pyridine was treated with Selectfluor™ (4.6 g, 13 mmol) and heated at 50 °C. After 22 h, the reaction mixture was cooled to ambient temperature, poured into 150 mL of half-saturated NaHCO3 solution, and extracted with 2 × EtOAc (total 400 mL). The combined organic phase was dried over sodium sulfate, filtered, and concentrated. The crude material was purified by silica gel chromatography (0 to 20% EtOAc / hexane) to give the title compound (580 mg, 23% yield) as an orange solid.

[0174] [Steps] [2]: Synthesis of 6-bromo-8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazo[1,2-a]pyridine A suspension of dried zinc powder (240 mg, 3.7 mmol) in THF (3.0 mL) was treated with 1,2-dibromoethane (30 mL, catalyst) and heated at 50 °C to obtain a mixture. Trimethylchlorosilane (30 mL, catalyst) was then added. After 15 min, the mixture was cooled to ambient temperature. Dried lithium chloride (170 mg, 3.9 mmol) was added, followed by dropwise addition of a solution of 1-(bromomethyl)-2,5-difluoro-3-toluene (480 mg, 2.2 mmol) in THF (2.0 mL) (Note: exothermic reaction). The mixture was stirred at ambient temperature for 1 h. Simultaneously, a slurry of 6,8-dibromo-3-fluoroimidazolo[1,2-a]pyridine (580 mg, 2.0 mmol) and Pd(PPh 3) 2Cl 2 (41 mg, 0.059 mmol) in THF (3.0 mL) was degassed with nitrogen. A freshly formed zincate solution was transferred to this slurry via syringe and rinsed with 2 × 0.5 mL THF to ensure complete transfer. The resulting mixture was stirred at ambient temperature for 1 h 20 min, followed by stirring at 40 °C for 4 h. After cooling to ambient temperature, the reaction mixture was quenched with 4 mL of saturated NH 4Cl solution. The organic layer was concentrated, diluted with CH₂Cl₂ (10 mL), and filtered through a diatomaceous earth bed. The concentrated filtrate yielded a brown residue, which was purified by silica gel chromatography (for the CH₂Cl₂-loaded compound dissolved in 0 to 10% EtOAc / hexane) to give the title compound (440 mg, 63% yield) as a yellow solid.

[0175] [Steps] [3]: Synthesis of 8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazo[1,2-a]pyridine-6-carboxynitrile A reaction mixture consisting of 6-bromo-8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazolo[1,2-a]pyridine (440 mmol, 1.2 mmol), zinc cyanide (100 mg, 0.87 mmol), Pd₂(dba)₃ (46 mg, 0.050 mmol), and 1,1'-bis(diphenylphosphino)ferrocene (dppf) (41 mg, 0.075 mmol) in anhydrous DMF (5.0 mL) was degassed with nitrogen and then heated at 90 °C for 6 h. The reaction mixture was cooled to ambient temperature and treated with CH₂Cl₂ (50 mL), water (40 mL), and 28% ammonium hydroxide solution (4.0 mL). The aqueous layer was extracted with CH₂Cl₂ (50 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to give a brown oily substance, which was purified by column chromatography (0 to 20% EtOAc / hexane gradient) to give the title compound (310 mg, 81% yield) as a light brown solid. LC / MS ES⁻¹ + m / z = 302.8 [M+H]⁺.

[0176] [Steps] [4]: Synthesis of 8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazo[1,2-a]pyridine-6-carbamate A solution of 0.50 N sodium methoxide in methanol (1.0 mL, 0.50 mmol) was added to a suspension of 8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazolo[1,2-a]pyridine-6-carboxynitrile (150 mg, 0.50 mmol) in methanol (6.0 mL). After stirring at ambient temperature for 4 h 30 min, ammonium chloride (270 mg, 5.0 mmol) was added and the reaction mixture was stirred for 18 h. The resulting mixture was concentrated under vacuum, treated with 10% NaHCO3 aqueous solution (10 mL) and subjected to sonication to obtain a suspension. After stirring for 1 h, the product was collected by filtration, washed with water (10 mL), and dried under vacuum to give the title compound (170 mg, >100% yield) as a light brown solid. It was used in the next step without further purification. LC / MS ES + m / z = 319.7 [M+H]+.

[0177] [Steps] [5]: Synthesis of 2-(8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazo[1,2-a]pyridine-6-fluoropyrimidin-4-ol Add (Z)-3-ethoxy-2-fluoro-3-sideoxyprop-1-en-1-ol sodium sodium (310 mg, 2.0 mmol) to a suspension of 8-(2,5-difluoro-4-methylbenzyl)-3-fluoroimidazole[1,2-a]pyridine-6-carbamate (0.50 mmol, derived from the theoretical amount in the previous step) in ethanol (5.0 mL). Heat the reaction mixture in a sealed vial at 90 °C for 16 h. After cooling to ambient temperature, dilute the mixture with water (7.5 mL) and adjust the pH to 4 with 1N HCl aqueous solution. The resulting light brown solid was collected by filtration, washed with water (50 mL) and ethyl ether (30 mL), and dried to give the title compound (140 mg, 71% yield, in two steps) as a brown solid. ¹H NMR (500 MHz, DMSO-d6) δ (ppm) 12.8 (br. s, 1 H), 8.98 (s, 1 H), 8.22 (br. s, 1 H), 7.74 (d, 1 H), 7.35 (br. s, 1 H), 7.15 (m, 1 H), 4.50 (s, 2 H), 2.18 (s, 3 H).

[0178] [] [Compound] [I-20] The title compound was synthesized in two steps:

[0179] [step] [1]: Synthesis of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate A solution of 25 wt% sodium methoxide in methanol (2.1 mL, 9.3 mmol) was added to a solution of 500 mg, 1.9 mmol, of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carboxynitrile (22 mL) in methanol. After stirring at ambient temperature for 1 h, ammonium chloride (1.0 g, 19 mmol) was added and the reaction mixture was stirred overnight. The reaction mixture was concentrated under vacuum, diluted with a semi-saturated NaHCO3 solution (20 mL) and a 1.0 N sodium hydroxide solution (2.0 mL), and extracted with 2 × 20 mL of EtOAc. The combined organic phases were dried over sodium sulfate, filtered, and concentrated to give a crude product as a brown solid. This crude product was used in the next step without further purification. LC / MS ES + m / z = 288.1 [M+H]+.

[0180] [step] [2]: Synthesis of 5-fluoro-2-(8-(2,5-difluorophenylmethyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol Add (Z)-3-ethoxy-2-fluoro-3-sideoxyprop-1-en-1-ol sodium salt to a suspension of 8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-carbamate (500 mg, 1.7 mmol) in ethanol (9.0 mL). Heat the reaction mixture in a sealed vial at 90 °C for 2 h. After cooling to ambient temperature, acidify the mixture to pH 4 by dropwise addition of concentrated HCl solution. Concentrate the resulting mixture under vacuum. Purify by preparative reversed-phase HPLC (acetonitrile-water gradient with 0.1% TFA as an additive) to give the title compound as a yellow solid (200 mg, 28% yield, in 2 steps). 1H NMR (500 MHz, DMSO- d 6) δ (ppm) 12.6 (br. s, 1 H), 9.49 (s, 1 H), 8.32 (s, 1 H), 8.19 (br. s, 1 H), 7.89 (s, 1 H), 7.43 (s, 1 H), 7.25 (m, 1 H), 7.13 (m, 1 H), 4.58 (s, 2 H).

[0181] [Compound] [I-20] [Of] [Na+] [salt] [] Under a nitrogen atmosphere, 5-fluoro-2-(8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-ol ( [Compound] [I-20] (10 g, 28 mmol) was added to a light brown suspension in 450 mL of anhydrous MeOH, followed by a solution of 0.50 N sodium methoxide in methanol (57 mL, 28 mmol). After simple sonication, the resulting light orange solution was stirred at ambient temperature for 15 min and concentrated to dryness under vacuum. The solid was resuspended in 200 mL of ether by sonication and concentrated (twice). The resulting solid was resuspended in 500 mL of ether and stirred at ambient temperature for 3 h. The solid was collected by vacuum filtration and washed with ether (3 × 100 mL). After drying on the filter overnight, the product salt was dried in a vacuum oven at 45 °C for 5 days to obtain sodium 5-fluoro-2-(8-(2,5-difluorobenzyl)imidazo[1,2-a]pyridine-6-yl)pyrimidin-4-olide (11 g, 99% yield) as a light brown solid. 1H NMR (500 MHz, D 2O) δ (ppm) 8.90 (s, 1 H), 7.98 (d, 1 H), 7.95 (d, 1 H), 7.70 (d, 1 H), 7.10 (m, 1 H), 6.98-6.89 (m, 2 H), 4.53 (s, 2 H).

[0182] [Biology Section] [, , ] [Mode] [I] [Evaluation of the biological properties of compounds] [, , ]

[0183] This invention also provides for the evaluation of the biological properties of compounds of Formula I. The activity of the compounds of this invention as representative sGC stimulators was tested in vitro in various cells and analyses, and their ability to lower blood pressure in animals was tested in vivo. Blood pressure reduction serves as an indicator of the compound's ability to bind to peripheral targets in vivo. Other tests were used as indicators of the compounds' ability to cross the BBB, bind to targets in the CNS, increase cGMP levels in the CNS, and thereby stimulate functional responses in animals. These biological properties represent another embodiment of the invention.

[0184] [Example] [2] [:] [Based on] [cGMP Glo Sensor] [Cellular Analysis] [,]

[0384] [hole] [Format for bioactivity measurement] [, , ] Human embryonic kidney cells (HEK293) containing GloSensor™ 40F cGMP (product number: CS182801, Promega) were used to evaluate the activity of test compounds. A luminescent biosensor (engineered luciferase) incorporated into these cells detects cGMP formed by compounds stimulating the sGC enzyme and emits fluorescence.

[0185] cGMP GloSensor cells were maintained in Dulbecco's Modification of Eagle's Medium (DMEM) supplemented with fetal bovine serum (FBS, 10% final) and hygromycin (200 μg / ml). The day before analysis, cells were seeded at a density of 1.5 × 10⁴ cells / well in 50 µL of 10% FBS DMEM in 384-well flat white-bottomed poly-D-lysine-coated dishes (Corning catalog 35661). Cells were cultured overnight at 37°C in a humidified chamber with 5% CO₂. The following day, the medium was removed and cells were replaced with 40 µL / well of GloSensor™, 2 mM (Promega, catalog E1291). Cells were treated at 25°C for 90 min to allow the matrix to equilibrate within the cells. The test compound and diethyltriamine NONO salt (DETA-NONO salt) were diluted to 3 mM (20×) in serum-free, CO2-independent medium and serially diluted with 4× diluent to generate a 5X dose profile. 10 μL was added to each well (x μM for the test compound solution and 10 μM for the DETA-NONO salt solution; where x is one of the following final concentrations: 30 μM, 7.5 μM, 1.9 μM, 469 nM, 117 nM, 29.3 nM, 7.3 nM, 1.83 nM, 0.46 nM, 0.11 nM, 0.03 nM) for kinetic studies. Fluorescence was immediately measured using an Envision (Perkin Elmer) at 0.2 sec / well. For endpoint SAR screening, data were collected after incubation at room temperature for 55 min.

[0186] Concentration-response data were analyzed using a 4-parameter fitting (logarithmic (promoter) and response-variable slope) method. EC 50 was derived from curve fitting and defined as the concentration at which a compound initiates 50% of its maximum response. When multiple experiments were performed on a given compound, the geometric mean of all experiments was reported.

[0187] Table A below summarizes the EC50 values ​​of the compounds of this invention in Globular analysis. Table A. [Compound Number] [Glo EC, 50 , (nM) ] [Compound Number] [Glo EC, 50 , (nM) ] I-21 B I-5 A I-2 B I-14* A I-1 B I-16 A I-4 B I-15 B I-23 B I-19 B I-20* A I-17 A I-11 B I-18 A I-13 B I-12 B I-10 B I-3 A I-6 A I-8 C I-7 B I-9 B sGC enzyme activity in HEK cells was determined using GloSensor analysis. The code for sGC enzyme activity (expressed as EC 50, defined as the concentration at which a compound induces 50% of its maximum response) is as follows: EC 50 ≤ 100 nM = A; 100 nM < EC 50 ≤ 1000 nM = B; 1000 nM < EC 50 = C. *For compounds I-20 and I-14, both the free acid and sodium salt were used, and the results here are the average of all experiments run independently of the form.

[0188] [] [Example] [3.] [Based on] [cGMP] [Analysis of neuronal cells for bioactivity measurement] Primary rat neurons were isolated from 18-day-old Spög-Dolly female fetuses. The fetuses were collected in Hank's balanced salt solution (HBSS) and rapidly removed from the brain. The hippocampus was isolated and mechanically fragmented. Further tissue digestion was performed at 37°C in Ca2+- and Mg2+-free HBSS with 0.25% (wt / vol) trypsin solution for 15 min. After trypsinization, the cells were washed and resuspended in neurotrophic medium supplemented with 0.5 mL M glutamic acid, 12.5 μM glutamic acid, 2% B-27, 100 U / mL penicillin, and 100 µg / mL streptomycin. Cells were seeded at a density of 26 × 10³, 3 × 10⁴, or 4 × 10⁴ cells / well in poly-D-lysine-coated 384-well flat transparent trays (Corning catalog number 354662). Cells were cultured at 37°C in a humidified chamber with 5% CO2 for 6–7 days. The culture medium was removed, and the cells were washed 1× with HBSS containing Ca2+ and Mg2+, and then replaced with 40 μL of HBSS containing 0.5 mM IBMX, and cultured at 37°C for 15 minutes. 10 μL of 5× test compound stock solution containing diethyltriamine NONO salt (DETA-NO) was added. The final concentration of DETA-NO was 10 μM or 30 μM. Cells were cultured at 37°C for 20 minutes. The culture medium was removed, 50 μL of ice-cold 10% acetic acid was added, and the cells were cultured at 4°C for 60 minutes. After centrifugation at 1000 × g for 5 minutes at 4°C to precipitate cell debris, the supernatant was aspirated into a clean dish, and the cGMP content of the samples was analyzed. The cGMP concentration of each sample was determined using LC-MS / MS.

[0189] Concentration-response data were analyzed using a 4-parameter fitting (logarithmic (promoter) and response-variable slope) method. EC 50 was derived from curve fitting and defined as the concentration at which a compound initiates 50% of its maximum response. When multiple experiments were performed on a given compound, the geometric mean of all experiments was reported.

[0190] Table B below summarizes the EC50 values ​​in neuronal analysis of the compounds of this invention. Table B. [Compound Number] [sGC] [Neuron] [EC, 50 , (nM) ] [Compound Number] [sGC] [Neuron] [EC, 50 , (nM) ] I-21 A I-3 A I-2 B I-15 B I-1 B I-9 A I-4 A I-19 A I-23 A I-5 A I-20* A I-14* A I-16 A I-12 A I-6 A I-7 A Cellular analysis based on neurons. EC 50 ≤ 100 nM = A; 100 nM < EC 50 ≤ 1000 nM = B; 1000 nM < EC 50 = C. *For compounds I-20 and I-14, both free acid and sodium salt were used, and the results here are the average of all experiments run independently of the form.

[0191] [] [Example] [4] [:] [right] [CHO-K1] [in cells] [Stablize] [Performance] [Human] [α2β1 sGC] [Assay of the biological activity of isoenzymes] [] The sGC stimulant was dissolved in DMSO to prepare a 10 mM solution and stored at -20°C. To achieve the desired test concentration, the stock solution was serially diluted in DMSO and then diluted to the appropriate concentration in analytical buffer.

[0192] CHO-K1 cells stably transfected with human α2β1 sGC isoenzymes (produced by GenScript from Ironwood) were cultured in F-12K medium (ATCC catalog number 30-2004) containing 10% fetal bovine serum, 4 µg / mL puromycin (Gibco catalog number A11138-03), and 0.4 mg / mL genimycin (Gibco catalog number 10131-027). For GC activity assays, cells were seeded at densities of 3 × 10⁴ cells / well or 15 × 10³ cells / well in 50 μL or 70 μL of medium in 384-well poly-D-lysine-coated flat-bottomed dishes (Fisher Scientific #08-774-311). Cells were cultured for 24 hours at 37°C in a humid chamber supplemented with 5% CO₂.

[0193] For each test concentration, the compound was diluted 100-fold in 100% DMSO to its final analytical concentration. Immediately prior to analysis, this solution was diluted 20-fold in HBSS containing calcium, magnesium, and 50 µM DETA-NONO salt (5 × final analytical concentration). The culture medium was removed, and the cells were washed once with 40 μL HBSS. The cells were then cultured at 37°C for 15 min in 40 μL of HBSS containing 0.5 mM IBMX. The cells were then added to a 10 μL sGC stimulator / HBSS / DETA-NONO salt dish and incubated at 37°C for another 20 min. The final DMSO concentration was 1%, the final DETA-NONO salt concentration was 10 µM, and the final compound concentrations were 30,000 nM, 6,000 nM, 1,200 nM, 240 nM, 48 nM, 9.6 nM, 1.92 nM, 0.384 nM, 0.077 nM, 0.015 nM, or 0.003 nM.

[0194] After incubation with the compound, remove the analysis buffer and add 50 μL of ice-cold 10% acetic acid + 150 ng / mL internal standard (+3 cGMP) to each well. Incubate the samples on ice for 30 to 60 min. Centrifuge at 1000 × g for 5 min at 4 °C to precipitate cell debris, transfer the supernatant to a clean dish, and analyze the cGMP content of the samples.

[0195] Data were analyzed using GraphPad Prism software v.8 with 4-parameter fitting (logarithmic (activator) and response-variable slope). EC 50 was derived from curve fitting and defined as the concentration at which a compound initiates 50% of its maximum response. When multiple experiments were performed on a given compound, the geometric mean of all experiments was reported.

[0196] Table C below summarizes the EC 50 values ​​of the compounds of this invention in CHO analysis. Table C [Compound Number] [sGC CHO] [EC, 50 , (nM) ] [Compound Number] [sGC CHO] [EC, 50 , (nM) ] I-21 A I-15 B I-2 B I-19 A I-17 A I-18 A I-4 A I-5 A I-23 B I-14* A I-20* A I-16 A CHO cell analysis. EC 50 ≤ 100 nM = A; 100 nM < EC 50 ≤ 1000 nM = B; 1000 nM < EC 50 = C. *For compounds I-20 and I-14, both the free acid and sodium salt were used, and the results here are the average of all experiments run independently of the form.

[0197] [Example] [5] [:] [After acute doses at multiple concentrations of representative compounds of the present invention] [,] [Effect on blood pressure in normotensive rats] [] [a)] [Compound] [I-14]

[0198] Male, normotensive Sporgodory rats were purchased from Charles River Laboratories. These rats had indwelling femoral artery catheters implanted. The animals were tethered to a tethering system connected to pressure sensors to monitor cardiovascular (CV) parameters, specifically mean arterial pressure (MAP) and heart rate (HR). The animals were acclimatized to the system overnight, and baseline CV parameters were collected. Subsequently, awake, freely moving rats were administered single oral doses of 1, 3, 10, and 30 mg / kg of [the drug / treatment] containing [the drug / treatment]. [Compound] [I]-

[14] Milli-Q water (a dose produced by the sodium salt of compound I-14). Blood samples were collected from each animal via catheter before and 2 hours after administration for compound concentration quantification. Hemodynamic measurements were recorded within 10 hours after administration. Fifty-four male rats were used for these studies and housed in a range of weights from 250 to 275 grams. They were housed individually in a controlled environment of temperature (21 ± 1°C) and relative humidity (36 ± 1%) in a 12-hour light-dark cycle room (lights on at 6:00 AM and off at 6:00 PM) at the SmartLabs housing (21 Erie Street, Cambridge, MA). Animals were allowed free access to food (LabDiet Prolab Isopro RMH 3000, St. Louis, MO). Two groups of studies were conducted. For the first group of studies, [Compound] [I] [-14] Reconstituted in Milli-Q water at 0.3 and 1.0 mg / ml and frozen at -20°C. For the second group of studies, weighing was performed at Cyclerion Therapeutics. [Compound] [I]-

[14] sodium salt and restored in SmartLabs to provide [Compound] [I]-

[14] 0.1, 0.3, 1.0 and 3.0 mg / ml solutions in Milli-Q water. The prepared formulations should be thawed and stored at room temperature within 4 hours before administration, or prepared and stored at room temperature within 2 hours before administration.

[0199] The study was conducted in six independent phases. The total number of individuals and the allocation of treatment are listed in the table below. Animals were initially given the treatment within 3 days of receipt, and if the catheter remained patent, it was administered again 6 to 7 days after clearance. Animals were not given the treatment more than twice. [Group] [Number of animals] [Therapeutic Agent] [Dose-volume] [Dose Concentration] 1 13 Mediator 10 mL / kg - 2 9 1 mg / kg [Compound] [I-14] 10 mL / kg 0.1 mg / mL 3 6 3 mg / kg [Compound] [I-14] 10 mL / kg 0.3 mg / mL 4 11 10 mg / kg [Compound] [I-14] 10 mL / kg 1.0 mg / mL 5 6 30 mg / kg [Compound] [I-14] 10 mL / kg 3.0 mg / mL

[0200] [, , ] [, Blood pressure measurement , ] [, , ] This study used conscious, freely moving rats fitted with an ADInstruments LabChart (v8) tether to a blood pressure sensor (Harvard Apparatus catalog number APT300) to collect hemodynamic data. After overnight acclimatization to the tether and pressure sensor, the animals were administered medication 1 hour after baseline recording. The dose volume was 10 mL / kg. [Compound] [I]-

[14] Sodium salt form or a single oral (PO) dose of the medium. Data collection within 10 hours after continued administration.

[0201] Hemodynamic data were monitored and output using ADInstruments LabChart (v8). Blood pressure and heart rate were continuously monitored, and data were collected at 1000 data points / second, then averaged into 10-minute groups for analysis. The change in MAP (ΔBMAP) and change in HR (ΔBHR) relative to baseline were calculated using Microsoft Excel 365 with averaged pre-dose baseline over a 1-hour period prior to administration. Peak ΔVMAP, time to peak ΔVMAP, peak ΔVHR, and time to peak ΔVHR were determined using this 10-minute grouped dataset. The datasets were further merged into 1-hour groups for MAP and HR plots and for analysis of ΔBMAP, ΔBMAP, and ΔBHR. Definitions of these terms / abbreviations are summarized below: ΔBMAP, dMAP Changes in mean arterial pressure relative to baseline Δ BHR, dHR Changes in heart rate relative to baseline Δ VMAP Mediator-adjusted mean arterial pressure Δ VMAP Mediated heart rate AOC Area on the curve

[0202] Statistical analysis was performed using GraphPad Prism (v8). The significance of ΔBMAP and ΔBHR compared to catalyzed rats was determined by a two-factor repeated-measures ANOVA followed by a Dunnett multiple comparison test. If missed data points existed, mixed-effects analysis was used. Catalyzed-adjusted MAP (ΔVMAP) was calculated by subtracting the ΔBMAP of the catalyzed group from the ΔBMAP of each dose group at each time point. Catalyzed-adjusted HR (ΔVHR) was calculated in a similar manner to ΔVMAP.

[0203] The significance of AOC data compared to the mediator was determined by using a one-way ANOVA followed by a Dunnett multiple comparison test.

[0204] Some data were removed prior to analysis. Data collected at 130 min and 140 min post-dose administration were removed due to 2-hour blood sample collection. Several time points of a rat at 1 mg / kg were excluded due to signal loss during the experiment, which began at 470 min and continued until the end of the study (600 min). For various reasons, the entire time series of five animals was excluded from all datasets, including outliers used for specific analyses or signal loss leading to abnormal results.

[0205] [, , ] [, Blood pressure changes , ] [, , ] The change in MAP relative to baseline (ΔBMAP) is graphically shown in Figure 1. Compared to rats treated with catalytic agents, using... [Compound] [I]-

[14] Treatment resulted in a greater reduction in MAP in rats (as assessed by change in MAP relative to baseline, ΔBMAP). The ΔBMAP dataset was significant using a two-factor ANOVA (p < 0.0001 for both treatment dose and time, and p = 0.045 for the treatment dose × time interaction). A Dunnett multiple comparison test of the primary therapeutic effect showed p = ns for 1 mg / kg, p = 0.0034 for 3 mg / kg, and p < 0.0001 for 10 and 30 mg / kg doses compared to mediator-treated rats. A single Dunnett multiple comparison test of the single effect compared to mediator-treated rats at various time points and doses showed ΔBMAP at 10 and 30 mg / kg doses. [Compound] [I]-

[14] The reduction was greater in treated rats over the entire 6-hour period following administration; at 3 mg / kg [Compound] [I]-

[14] The reduction was greater at 1-hour, 2-hour, and 3-hour post-administration in treated rats; while in rats treated with 1 mg / kg [Compound] [I]-

[14] No reduction was observed in the treated rats.

[0206] Calculations were made using datasets grouped in 10-minute intervals. [Compound] [I-14] The maximum effect on ΔVMAP is shown in Table D below: Table D. [Compound] [I-14] The maximum effect of mediator-adjusted MAP (ΔVMAP) [Compound] [I-14] [peak] [Δ, V MAP (mm Hg) [Time of arrival at the peak] [(min)] 1 mg / kg -6.2 90 3 mg / kg -10.1 70 10 mg / kg -22.3 30 30 mg / kg -24.4 30

[0207] Based on main effect analysis, single effect analysis, and AOC assessment, the dose with no effect on ΔBMAP was 1 mg / kg.

[0208] [, in conclusion , ] [, , ] Relative to the baseline and adjusted for self-propelled agents, [Compound] [I]-

[14] MAP was reduced at 3, 10 and 30 mg / kg.

[0209] [b)] [Compound] [I] [-20] [] use [Compound] [I]-

[20] Similar studies to those described above were conducted. Awake, freely moving rats were administered single oral doses of 1, 3, 10, and 30 mg / kg of [a specific ingredient / product]. [Compound] [I]-

[20] (from compound I) [-20] sodium salt (prepared at a dose) in Milli-Q water. Blood samples were collected from each animal via catheter before and 2 hours after administration for compound concentration quantification. Hemodynamic measurements were recorded over ten hours after administration.

[0210] [] [, Blood pressure changes , ] [, , ] The change in MAP relative to baseline (ΔBMAP) is graphically shown in Figure 2. Compared to rats treated with a mediator, using [Compound] [I]-

[20] Treatment resulted in a greater reduction in MAP in rats (as assessed by change in ΔBMAP relative to baseline MAP). ΔBMAP was significant in the two-factor ANOVA dataset (p < 0.0001 for the treatment agent and treatment agent × time interaction; and p = 0.022 for time). A Dunnett multiple comparison test of the primary therapeutic effect showed p = 0.055 (ns) for 1 mg / kg, p = 0.0001 for 3 mg / kg, and p < 0.0001 for 10 and 30 mg / kg doses compared to the cartel-treated rats. A single Dunnett multiple comparison test of the single effect compared to the cartel-treated rats at each time point and dose showed MAP at 3, 10, and 30 mg / kg. [Compound] [I]-

[20] The reduction was greater in treated rats over the entire 6-hour period following administration; at 1 mg / kg [Compound] [I]-

[20] The reduction was greater at 1-hour, 2-hour, 3-hour, 4-hour and 5-hour post-administration in treated rats. []

[0211] Calculations were made using datasets grouped in 10-minute intervals. [Compound] [I-20] The maximum effect on ΔVMAP (MAP adjusted by the agent) is shown in Table E below. Table E. [Compound] [I] [-20] The maximum effect of mediator-adjusted MAP (ΔVMAP) [Compound] [I-20] [peak] [Δ, V MAP (mm Hg) [Time of arrival at the peak] [(min)] 1 mg / kg -14.3 20 3 mg / kg -19.2 20 10 mg / kg -20.2 30 30 mg / kg -38.1 30

[0212] [, , ] [, in conclusion , ] [, , ] [Compound] [I]-

[20] MAP was reduced at 1, 3, 10 and 30 mg / kg relative to baseline and as adjusted for self-mediating agents.

[0213] [] [c)] [other] [BP] [Measurement] [] In studies similar to those described above, the drug was formulated with PEG400 and administered at a dose of 10 mg / kg. [Compound] [I]- [4] The maximum reduction in MAP relative to baseline (ΔBMAP) was 20 mm Hg at 50 min after administration. In studies similar to those described above, the drug was prepared in PEG400 and administered at 10 mg / kg. [Compound] [I]-

[20] The peak ΔBMAP was -26 mm Hg at 42 min after administration. [Compound] [I]-

[20] In another study, tested at 1, 3 or 10 mg / kg and formulated in methylcellulose, the compound reduced MAP from baseline at all tested doses.

[0214] [Example] [6] [:] [In primary neurons of rats] [sGC] [Induced by stimulants] [CREB] [Phosphorylation] [, , ] [, Target , ] To evaluate the ability of the compounds of this invention to activate cAMP response element-binding protein (CREB) in primary rat neurons. CREB is a cellular transcription factor. It binds to a DNA sequence called a cAMP response element (CRE) and regulates the transcription of downstream genes (see Bourtchuladze R, et al., Cell 1994;79(1): 59-68). CREB has a well-documented role in neuronal plasticity and long-term memory formation in the brain, and has been shown to be indispensable in spatial memory formation (see Silva AJ, et al., Annual Review of Neuroscience 1998;21: 127-148). CREB protein is activated by phosphorylation of serine 133 by various kinases, including cAMP-dependent protein kinase or protein kinase A (PKA), cGMP-dependent protein kinase or protein kinase G (PKG), and Ca2+ / calmodulin-dependent protein kinase. (See Shaywitz AJ and Greenberg ME, Annual Review of Biochemistry 1999;68(1): 821–861 and Wong JC et al., J Cell Biochem 2012: 113(11):3587-98). Stimulation of CREB may have therapeutic benefits for diseases involving cognitive, neuronal plasticity, and / or neuronal dysfunction.

[0215] [, , ] [, primary rat neuron culture , ] [, , ] Neurons were isolated from Spögdolly rat embryos on day 18 (E18). Approximately 10 embryos were obtained from each rat, and the entire brain was isolated from these embryos. The hippocampus and cortex were dissected from the brain using two fine forceps under a stereomicroscope. The meninges were carefully removed. After dissection, the tissue was minced and gently washed once in a 15-mL conical tube with 10 mL of Hank's balanced salt solution (HBSS, Corning catalog number 21-022-CM) free of Ca2+ and Mg2+. After washing, 5 mL of a solution of 0.25% trypsin (Invitrogen catalog number 15090-046) and 0.1% deoxyribonuclease I (DNase I, Sigma catalog number DN-25) was added to the tissue, followed by incubation at 37°C for 15 min. Next, the tissue was washed three times with ice-cold HBSS, and 3 mL of 0.1% DNase I solution was added. The tissue was then slowly aspirated 12 times using a glass Pasteur pipette, followed by centrifugation at 500 × g for 10 min. The cell pellet was resuspended in culture medium (Neuro-Matrix Medium, Gibco catalog number 21103-049), 2% B27 supplement (Gibco catalog number 17504-044), 0.5 mM L-glutamic acid (Corning catalog number 25-005-Cl), 25 µM L-glutamic acid (Sigma catalog number G1251), and 1% penicillin / streptomycin (Gibco catalog number 15070-063). Subsequently, the cell suspension was seeded at 100,000 cells / well in poly-L-lysine-coated 96-well plates. Twenty-four hours after inoculation, the general culture medium was removed and replaced with the medium described above but without glutamate. Cells were maintained in a humid incubator at 37°C with 5% CO2 and used for analysis between days 6 and 10 post-collection.

[0216] [, Analysis conditions , ] [, , ] For each test concentration, the compound was diluted to 100× its final analytical concentration in 100% DMSO. Immediately before analysis, the compound was diluted 1 / 10 in HBSS (containing calcium and magnesium) (10× final analytical concentration) containing 100 µM DETA-NONO salt (10× final analytical concentration). The culture medium was removed, and the cells were washed once with 90 μL HBSS (Corning Catalogue No. 21-023-CV). The cells were then cultured at 37°C with 90 μL HBSS for 30 min. Cells were added to a 10 μL test compound / HBSS / DETA-NONO salt culture dish and cultured at 37°C for another 30 min. The final DMSO concentration was 1%, the final DETA-NONO salt concentration was 10 µM, and the final compound concentrations were 10 µM, 1 µM, 0.1 µM, 0.01 µM, 0.001 µM, 0.0001 µM, 0.00001 µM, and 0.0 µM. The culture medium was removed, cells were lysed, and pCREB content was determined according to the Cisbio protocol (phosphorylated CREB (Ser133) catalog number 64CREPEG). Culture discs were read using an Envision instrument (PerkinElmer).

[0217] [, Data Analysis , ] [, , ] pCREB was measured in each well and analyzed using GraphPad Prism software v.8 with a 3-parameter fit (logarithm (activator) and response-variable slope). EC50 was extrapolated from the curve fit and defined as the concentration at which the sGC stimulator compound induced its maximum response. Multiple experiments were performed on the given compounds, and the geometric mean of all experiments was reported. For [Compound]

[14] and

[20] Both use free acid in these experiments.

[0218] Table F below summarizes the EC50 values ​​of the compounds of this invention in pCREB analysis. Table F. [Compound Number] [pCREB] [EC, 50 , (nM) ] [I-14] twenty one [I-20] 13

[0219] [] [Example] [7] [:] [Rat cerebrospinal fluid] [(CSF)] [Pharmacokinetic Properties] [, , ] [, plan , ] [, , ] PK was determined in rats after oral administration. For the oral (PO) test, a group of six male Spög-Dolly rats with an indwelling catheter placed in the cerebellomedullary cistern were used. The PO group was administered 3.0 or 10 mg / kg of the compound in a solution of PEG400 or a suspension in 0.5% Tween 80 and 0.5% methylcellulose / water. The PO dose was administered orally via a feeding tube and delivered to the stomach using a syringe and feeding tube. After oral administration, the feeding tube was flushed with approximately 0.5 mL of water to ensure complete delivery of the full dose. [, , ]

[0220] Plasma samples were collected as follows: CSF and blood samples were collected at 1 hour, 2 hours, and, if necessary, 4 hours after administration. CSF samples (0.05 mL) were collected via an intracisional catheter. Blood samples (0.25 mL) were collected via a tail incision. These samples were kept on ice until plasma processing. Blood samples were centrifuged at approximately 5°C and 3200 rpm for 5 minutes within 1 hour of collection. Plasma was directly transferred to individual Eppendorf tubes (0.125 mL). A stopper was placed on the catheter, and the catheter was frozen at approximately -70°C and stored until analysis. Plasma and CSF were collected, and the presence of compounds was analyzed.

[0221] [, Quantitative analysis of compounds. , ] [, , ] The compounds and internal standards under discussion were extracted from plasma by precipitation and from CSF by precipitation or dilution. Samples were analyzed using liquid chromatography (LC) and tandem mass spectrometry detection (MS / MS) with electrospray ionization. The standard curve ranged from 0.1 to 1000 ng / mL. The results for the compounds described herein in this analysis are shown in Table G (for a 10 mg / kg dose and / or 3 mg / kg of the compound). The compound concentrations from several animals were combined to obtain the geometric mean at each specific dose and time point.

[0222] Kp,uu is defined as the ratio of the concentration of unbound drug in CSF to the concentration of unbound drug in plasma. The unbound drug in plasma (or free plasma concentration) was calculated by multiplying the total plasma concentration by the unbound fraction as determined by plasma protein binding. Then, the CSF concentration was divided by the free plasma concentration to determine Kp,uu. (See, for example, Di et al., J. Med. Chem., 56, 2 - 12 (2013)) Table G. [Compound number] [Dose (mg / kg)] [1 h] [Kp,uu below] [2 h] [Kp, uu below] [4 h] [Kp, uu below] I - 20 10 B C C I - 1 3 A B A I - 4 3 A B B I - 20 3 C C C I - 5 3 C C C I-14 3 D D D I-16 3 D D D I-14 10 D D C I-18 3 B B B Rat CSF PK. Kp,uu ≤ 1 = A; 1 < Kp,uu ≤ 2 = B; 2 < Kp,uu ≤ 3 = C, 3 < Kp,uu = D.

[0223] [, , ] [Example] [8] [:] [Microdialysis experiment on rat brain] [] [, Target , ] [, , ] The objective of this study was to evaluate the levels of the sGC stimulant of this invention in the interstitial fluid (ISF) of the hippocampus and striatum, as well as in circulating plasma, after administration to male Spöghold's rats. To this end, microdialysis probes were implanted into the hippocampus, striatum, and jugular vein (JVC) cannulas in the rats. After collecting one pre-administration ISF sample, the animals were administered the sGC stimulant. Following administration, samples were collected via the hippocampal and striatum probes for 24 hours, and continuous plasma samples were collected via the JVC. All collected samples were stored at -80°C pending analysis of the compound content in the permeate.

[0224] [] [, Materials and Methods , ] [, , ] [animal] [] Five Spögdolly rats with pre-inserted jugular vein cannulas were used in this study. Upon arrival, the rats were grouped and housed in polycarbonate cages (2 to 3 rats / cage) and allowed to acclimatize for at least 3 days before the start of the study. The animals were housed in environments maintained at 22°C. Rats were kept at room temperature of [+]2°C and approximately 50% humidity for 12 hours under light / dark cycles with free access to food and water. Rats were tracked using unique identification numbers. Experiments were conducted according to protocols approved by the Institutional Animal Care and Use Committee of Charles River Laboratories South San Francisco.

[0225] [, Preparation and administration , ] [] On the day of treatment, freshly prepared sGC stimulant was administered to the animals as follows: [Compound] [I]-

[14] It was administered at a dose of 3 mg / kg in its sodium salt form at a concentration of 3 mg / mL in MilliQ water.

[0226] [In vitro experiments] [] In vitro Metaquant microdialysis (MQ-MD) experiments were performed to assess compound recoveries via the probe membrane. For this purpose, MQ probes (polyacrylonitrile, 3 mm membrane) were connected to a microperfusion pump (Harvard PHD 2000 syringe pump, Holliston, MA, or similar) via an inlet PEEK tubing. The probes were individually placed in an artificial CSF (aCSF) + 0.2% bovine serum albumin (BSA) bath containing 50 ng / mL sGC stimulant. The bath contents were continuously stirred and maintained at 37°C. Each probe was perfused with aCSF + 0.2% β-cyclodextrin (β-CD, slow flow) and carried by ultrapure water + 0.2% BSA. The flow rate was 0.15 µL / min for slow flow and 0.8 µL / min for carry flow. The probe outlet was connected to an automated fraction collector (820 microsampler, Univentor, Malta, or similar) via an outlet PEEK tubing. After perfusion stabilization, samples were collected into polypropylene vials over a 20-minute period. In individual vials, 150 µL of bath contents were collected at the beginning and end of the sample collection phase. The compound content of the in vitro dialysis fluid and bath samples was analyzed. Probe recovery was calculated as the ratio of the final compound concentration in the permeate sample to the bath concentration and expressed as a percentage recovery.

[0227] [Microdialysis procedure] [] Rats were anesthetized with isoflurane (2%, 800 mL / min O₂). Local anesthesia was administered with bupivacaine, and preoperative / postoperative analgesia was provided with carprofen. Animals were placed in a stereotactic frame (Kopf instrument, USA). Next, a MetaQuant microdialysis probe (polyacrylonitrile; 3 mm exposed membrane) was implanted into the striatum (STR) and hippocampus (HIPP). The probe tip coordinates for the STR were: anterior-posterior (AP) distance from the anterior fontanelle = +0.9 mm, lateral (L) distance from the midline = +3.0 mm, and ventral (V) distance from the dura mater = -7.0 mm, with the rack set at -3.3 mm. A second probe was then implanted into the hippocampus (HIPP). The coordinates of the probe tip for HIPP were: anterior-posterior (AP) distance from the anterior fontanelle = -5.3 mm, lateral (L) distance from the midline = -4.8 mm, and ventral (V) distance from the dura mater = -7.0 mm; the rack was set to -3.3 mm. After surgery, the animal was kept in a cage alone and provided with food and water at will.

[0228] Microdialysis experiments were performed one day post-surgery. The microdialysis probe was connected to a microinfusion pump (Harvard PHD 2000 syringe pump, Holliston MA) using a flexible PEEK catheter. The microdialysis probe was perfused with aCSF containing 147 mM NaCl, 3.0 mM KCl, 1.2 mM CaCl2, 1.2 mM MgCl2, and 0.2% β-cyclodextrin at a slow flow rate of 0.15 μL / min and H2O + 0.2% BSA at a rate of 0.8 μL / min. The microdialysis sample was collected over a 30-minute period using an automated fraction collector (820 microsampler, Univentor, Malta) into 300 μL polypropylene vials containing 15 µL of 0.02 M formic acid + 0.04% ascorbic acid / ultrapure water. After stabilization, a baseline sample was collected, and sGC stimulant was administered orally (PO) at T=0, with continuous sample collection for 24 hours (collected at 1h, 2h, 3h, 4h, 5h, 6h, 8h, 12h, and 24h). Samples were aliquoted for sGC stimulant analysis. All ISF samples were stored at -80°C until analysis.

[0229] In addition to ISF collection, blood samples were collected via JVC into K2+EDTA vials at T = -0.5 h, 0.5 h, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 12 h, and 24 h post-treatment. Blood was stored on ice until plasma processing (centrifugation at 4°C, 2,500 g for 10 min). Plasma was aliquoted into 1.5 mL Ependorf vials and stored at -80°C for analysis.

[0230] [Collection by the Post-Death Organization] [] After microdialysis, the animals were euthanized by CO2 asphyxiation. The brains were collected in 10% neutral buffered formalin for probe placement verification.

[0231] [, , ] [, Bioanalytical methods , ] [, , ] [Measuring the content of compounds in osmotic fluid and plasma] [] In osmotic fluid and plasma samples [Compound] [I]-

[14] The concentration was detected by ultra-high performance liquid chromatography (UPLC) combined with tandem mass spectrometry (MS / MS) and quantified in multiple reaction monitoring mode (MRM).

[0232] First, the plasma sample was mixed with an acetonitrile solution containing 100 ng / mL dexamethasone (internal standard) for protein precipitation. After incubation at room temperature for 5 minutes, the sample was centrifuged for 5 minutes (1300 rpm, 4°C), and the supernatant was diluted 100-fold in ultrapure water containing 0.1% formic acid.

[0233] Prior to analysis, an undiluted permeate sample (10 µL) was mixed with a 4 µL solution of internal standard containing 50 ng / mL dexamethasone (internal standard) dissolved in acetonitrile / ultrapure water (1:1) and 0.1% formic acid.

[0234] Undiluted permeate samples and diluted plasma supernatant were injected into a Shimadzu system (Shimadzu, USA) using an automated sample injector (Shimadzu Sil-30AC autosampler, Shimadzu, USA). Analytes were separated by liquid chromatography using a linear gradient of mobile phase B at a flow rate of 0.800 mL / min on a reverse-phase XBridge BEH C8 column (2.1 x 50 mm, 2.5 μm particle size; Waters, USA) maintained at 35 °C. Mobile phase A consisted of ultrapure water containing 0.1% formic acid. Mobile phase B consisted of acetonitrile containing 0.1% formic acid.

[0235] Data acquisition was performed in positive ionization mode using a QTrap® 5500 mass spectrometer (Applied Biosystems, USA) equipped with a turbine ion spray interface. The ion spray voltage was set to 5.5 kV and the probe temperature to 600°C. Collision gas (nitrogen) pressure was maintained at the culture medium. Quantification was performed using the following MRM transition range: m / z 372.0 / 352.0. A suitable introductory calibration curve was fitted using weighted (1 / x) regression. Sample concentrations were determined using these calibration curves. Accuracy was validated using quality control samples after each sample series. Calibration and quantification data were performed using the Analyst™ data system (Applied Biosystems, version 1.5.2).

[0236] [, Data Evaluation , ] [, , ] Data were plotted using Prism 8 on Windows (GraphPad Software, Inc.). The permeate concentration of the reported compound I-14 was corrected based on probe recovery. The average recovery was 14.7% (0.59 on SEM). [, result , ] [, , ]

[0237] All five animals were successfully administered the drug and completed the experiment. JVC was blocked in one animal at the 4-hour time point without collecting additional blood. No significant side effects were observed.

[0238] [In the striatum and hippocampus] [ISF] [China Investment Corporation and Compound Chemicals] [I] [-14] [Its function] [;] [STI / HIPP] [and] [Ratio of compounds in blood plasma] [] Figure 3 shows the STR and HIPP fractions of ISF osmotic fluid in adult male Sporg-Dolly rats after administration of (3 mg / kg; PO) compound I-14 at T = 0 min. [Compound] [I]-

[14] content.

[0239] Microdialysis allows sampling of compound concentrations in interstitial fluids between brain tissues. Given that tissues at sampling sites have different blood perfusion rates, physiological compositions, and clearance mechanisms compared to cerebrospinal fluid (CSF), we expect differences in distribution and measured concentrations between microdialysis studies and CSF-PK studies in the same animals. These differences will be observed in the time to reach maximum concentration, measured concentration, and ratios calculated from measured concentrations (Nagaya Y, Nozaki Y, Takenaka O, et al., Investigation of utility of cerebrospinal fluid drug concentration as a surrogate for interstitial fluid concentration using microdialysis coupled with cisternal cerebrospinal fluid sampling in wild-type and Mdr1a(- / -) rats. Drug Metab Pharmacokinet. 2016;31(1):57-66).

[0240] [, , ] [Example] [9] [:] [Single throw] [sGC] [Cephalospinal fluid of rats after stimulation] [(CSF)] [in] [cGMP] [Concentration Change] [(CSF)] [Biomarker Measurement] [) , , ] This experiment was conducted to determine the effect of different doses of the sGC stimulating compound of the present invention on the cGMP content in rat CSF. [, plan , ]

[0241] Rats were administered a single dose of the catalyst or sGC stimulant (1, 3, 10, or 30 mg / kg). CSF samples were collected and analyzed at one, two, and six hours after administration to determine cGMP and compound concentrations, and plasma samples were also collected and analyzed to determine compound concentrations. Each rat was sampled once or multiple times, with intervals of 3 days or more between administrations. Rats were fasted overnight the day before the experiment but allowed free access to water.

[0242] On the day of the experiment, the concentrations of compounds and cyclic guanosine monophosphate (cGMP) in the rat CSF were measured after oral administration. Male CD rats (250 to 275 g) with intracranial cannulas were used for these studies. Following protocol MIL-110, the rats were housed individually in a controlled environment of 21 ± 1°C and 36 ± 1% relative humidity in a SmartLabs housing (21 Erie Street, Cambridge, MA) with a 12-hour light-dark cycle (lights on at 6:00 AM and off at 6:00 PM). The animals were allowed free access to food (LabDiet Prolab Isopro RMH 3000, St. Louis, MO). Rats were administered the compounds of the present invention in the form of a suspension in 0.5% methylcellulose, 0.5% Tween 80, or MilliQ water at doses of 0 mg / kg (mediator), 1 mg / kg, 3 mg / kg, 10 mg / kg, or 30 mg / kg. The formulations were prepared, frozen at -20°C and thawed at room temperature for 1 hour prior to administration, or prepared and used within 2 hours, or prepared one day prior to administration and kept at room temperature with stirring overnight until administration. The oral dose was administered orally via a feeding tube into the stomach using a syringe and feeding tube. After oral administration, the feeding tube was flushed with approximately 0.5 mL of water to ensure complete delivery of the full dose.

[0243] Plasma and CSF samples were collected under isoflurane anesthesia as follows: CSF and blood samples were collected at 1, 2, and 6 hours after administration. CSF samples were collected via an intracisional catheter. Approximately 20 µL of CSF was collected and discarded (this includes a syringe filler volume of 14 to 16 µL); approximately 50 µL of CSF was then aspirated into an Ependorf tube containing 5 µL of glacial acetic acid. The CSF sample was rapidly frozen by immersion in liquid nitrogen. Next, while the animal remained anesthetized, blood samples were obtained via a tail incision and stored in K-EDTA tubes. These samples were kept on ice until plasma processing. Blood samples were centrifuged at approximately 5°C for 10 minutes at 3200 rpm within 1 hour of collection. Plasma was directly transferred to 96-well coils (0.125 mL). A stopper was placed on the catheter, and the catheter was frozen at approximately -70°C and stored until analysis. Plasma and CSF were collected, and the presence of compounds was analyzed.

[0244] [, Compounds and , ] [, cGMP , ] [, The quantitative measure. , ] The compounds, cGMP, and internal standards of this invention were extracted from plasma and CSF by precipitation. Samples were analyzed using electrospray ionization with liquid chromatography (LC) and tandem mass spectrometry (MS / MS). Standard curves for the compounds ranged from 0.1 to 1000 ng / mL. Standard curves for the cGMP ranged from 0.01 to 40 ng / mL. CSF cGMP data were plotted using GraphPad Prism, version 8.4.3, and expressed as mean ± SEM. Data were analyzed using mixed-effects analysis, followed by Dunnett's multiple comparison test for comparison with causative-treated rats at the same time points. Significance was set at p < 0.05.

[0245] [, , ] [, result , ] [, , ] One hour and two hours after administration, compared with rats treated with cartonants, rats administered 1, 3, and 10 mg / kg... [Compound] [I]-

[20] Rats treated with (in suspension form in Tween / MC) showed no significant change in cGMP concentration in rat CSF. However, six hours after administration, compared with rats treated with the carboxylate, rats treated with 10 mg / kg showed a significantly lower cGMP concentration. [Compound] [I]-

[20] The treated rats had significantly higher levels of cGMP in CSF. (See Figure 4)

[0246] Compared with rats treated with catalysts, rats treated with catalysts showed better results. [Compound] [I]-

[14] Rats treated with sodium salt had higher cGMP levels in CSF at all tested doses but not at all tested time points. 3 mg / kg was administered one and two hours after administration. [Compound] [I]-

[14] The rats had significantly higher cGMP concentrations in CSF. Two hours after administration, compared with rats treated with a carton, the cGMP concentrations were significantly higher in rats treated with 1 mg / kg CSF. [Compound] [I]-

[14] The treated rats had significantly higher levels of cGMP in CSF. At 1, 2, and 6 hours after administration, rats were given 10 or 30 mg / kg of the drug. [Compound] [I]-

[14] rats had significantly higher cGMP levels in CSF (see Figure 5).

[0247] [Example] [10A] [:] [Non-human primate cerebrospinal fluid] [(CSF)] [Pharmacokinetic Properties] [(] [Research] [A)] [, plan. , ] [, , ] The pharmacokinetic (PK) levels in NHP were determined after oral administration (PO). A group of four female cynomolgus monkeys was used to study the compounds. [Compound] [I]-

[20] The sodium salt was prepared into a 0.06 mg / mL solution in MilliQ water. [Compound] [I]-

[14] The sodium salt was prepared as a 0.2 mg / mL solution in MilliQ water. The preparation was transported frozen on dry ice, then thawed and thoroughly mixed before administration. A PO dose of 1 mg / kg was administered orally via tube feeding. [Compound] [I]-

[14] and 0.3 mg / kg [Compound] [I]-

[20] [。]

[0248] Plasma and CSF samples were collected as follows: CSF samples were collected 3 and 24 hours after PO administration. CSF samples (0.125 mL) were collected from the cerebellomedullary cistern via direct needle puncture with direct dilution. Blood samples (0.8 mL) were collected from peripheral veins at 0, 0.25, 0.5, 1, 2, 3, 6, 8, 12, 24, 32, and 48 hours. These samples were kept on ice until plasma processing. Proteins in the blood samples were precipitated using acetonitrile. Plasma was directly transferred to individual catheters (0.125 mL) using K₂EDTA as an anticoagulant. A cap was placed on the catheter, and the catheter was frozen at approximately -70°C and stored until analysis. Plasma and CSF were collected, and the presence of compounds was analyzed.

[0249] [, , ] [, Quantitative analysis of compounds. , ] [, , ] Plasma and CSF samples were analyzed using positive electrospray ionization with liquid chromatography (LC) and tandem mass spectrometry (MS / MS). The standard curve ranged from 0.1 to 1000 ng / mL.

[0250] The geometric mean of the equivalent values ​​obtained from the four animals in the studies of each compound were the concentrations in CSF and plasma, respectively.

[0251] Kp,uu is defined as the ratio of the concentration of unbound drug in the CSF to the concentration of unbound drug in plasma. The unbound drug in plasma (or free plasma concentration) is calculated by multiplying the total plasma concentration by the unbound fraction as determined by plasma protein binding. Then, the CSF concentration is divided by the free plasma concentration to determine Kp,uu. (See, for example, Di et al., J. Med. Chem., 56, 2-12 (2013))

[0252] The results for the compounds of the present invention are summarized in Table H below. Table H. [Compound number] [Dose (mg / kg)] [3 h] [Kp,uu below] [24 h] [Kp, uu below] I-20 0.3 A < / B I-14 1 C D NHP CSF PK. Kp,uu ≤ 1 = A; 1 < Kp,uu ≤ 2 = B; 2 < Kp,uu ≤ 3 = C; 3 < Kp,uu = D.

[0253] [Example] [10B] [:] [Non-human primate cerebrospinal fluid] [(CSF)] [Pharmacokinetic properties] [(] [Study] [B)] [, , ] The objective of this study was to investigate the [Compound] [I]-

[14] Pharmacokinetics and evaluation of the differences in pharmacokinetic values ​​when comparing CSF collected at the cerebellomedullary cistern (as in Example 10A) and the lumbar spine location. [, Research Design , ] [, , ] [Group ID] [Number of animals, a , ] [Test materials, , ] [Dosage level (mg / kg)] [Dose concentration (mg / mL)] [Dose-volume] [(mL)] [Route of administration / ] [plan] [collect] 1 4 F [Compound] [I-14] Total 1 0.1 / 0.1 5 On day 1, the single oral feeding dose of each test substance ● 15 and 30 min after administration; 1, 2, 3, 6, 8, 12, 24, 32 and 48 hours after administration [blood] ● 3 and 24 hours after administration [CSF] (CSF collected directly via the cerebellomedullary cistern) 2 4 F Total 0.3 0.03 / 0.03 5 On day 1, a single IV bolus injection of each test substance was performed. 3 4 F Total 1 0.1 / 0.1 5 On day 1, the single oral feeding dose of each test substance ● 15 and 30 min after administration; 1, 2, 3, 6, 8, 12, 24, 32 and 48 hours after administration [blood] ● 3 and 24 hours after administration [CSF] (CSF collected via the lumbar septum) 4 4 F Total 0.3 0.03 / 0.03 5 On day 1, a single IV bolus injection of each test substance was performed. No. = animal number; F = female; CSF = cerebrospinal fluid; No = number a, all groups will use the same animals, and there will be at least 7 days between the administration dates of each group. [, Test System , ] [, / , ] [, method , ] [, , ] Species: Long-tailed macaque (Macaca fascicularis) Species: Cynomolgus macaque Number of females: 4 Age: Adult Weight: 2.5 - 4 kg

[0254] Animals shall be housed in stainless steel cages equipped with stainless steel mesh bottoms and automatic water valves. The primary enclosure shall be as described in the Guidelines for the Care and Use of Laboratory Animals (National Research Council (NRC). Guide for the Care and Use of Laboratory Animals. Washington, DC: National Academy Press, 8th edition. 2011; Office of Laboratory Animal Welfare. Public Health Services Policy on Humane Care and Use of Laboratory Animals. Bethesda, Maryland: National Institutes of Health, revised 2015). These captivity conditions shall be maintained unless deemed inappropriate by the principal investigator and / or clinical veterinarian. Animals shall be socialized (where possible), except for times when separated for designated research procedures / activities.

[0255] In a crossover design, a single dose of compound I-14 was administered intravenously as a 0.15 mg / kg bolus solution and orally via tube feeding to four cynomolgus macaques. Plasma samples were collected at 0.25 (15 min), 0.5 (30 min), 1, 2, 3, 6, 8, 12, 24, 32, and 48 h after the IV and PO doses. CSF samples were collected from the cerebellomedullary cistern in groups 1 and 2 and from the lumbar vertebrae in groups 3 and 4 for all animals at 3 and 24 h. Sample extracts were prepared by protein precipitation, and compound I-14 concentrations were measured using LC-MS / MS. Pharmacokinetic parameters were calculated.

[0256] [Summarize] [, / , ] [in conclusion] [, , ] Overall, the mean pharmacokinetic parameters between IV (groups 2 and 4) and PO (groups 1 and 3) were within each other's standard deviation.

[0257] Compound I-14 in plasma was observed 3 and 24 hours after IV and PO administration using both cerebellomedullary cistern and lumbar spine sampling. At 3 hours after IV and oral administration, the geometric mean ratio of plasma CSF concentration to unbound concentration decreased within range C as described above, i.e., between 2 and 3, after both cerebellomedullary cistern and lumbar spine sampling (NHP CSF PK. Kp,uu ≤ 1 = A; 1 < Kp,uu ≤ 2 = B; 2 < Kp,uu ≤ 3 = C; 3 < Kp,uu = D). At 24 hours after IV and oral administration, the geometric mean ratio of plasma CSF concentration to unbound concentration decreased within ranges B and C after cerebellomedullary cistern sampling, and also within ranges B and C after lumbar spine sampling.

[0258] [Example]

[11] [:] [New Object Recognition with Enhanced Memory] [(NOR)] [Evaluation of the compounds of this invention in the model] [, , ] [, Target , ] To evaluate the efficacy of the CNS penetrant sGC stimulant of the present invention in reversing memory impairment induced by MK-801, a novel object recognition (NOR) test was used in male Long Evans rats.

[0259] [, introduce , ] NOR (Novel Oriented Recognition) is a test for recognizing learning and memory retrieval, utilizing the spontaneous preferences of rodents to study novel objects compared to familiar ones. NOR tests have been widely used to assess the potential of novel test compounds to enhance recognition properties. Because the NOR paradigm does not involve reward or harmful stimuli, it provides fewer confounding variables when translated into similar tests for human clinical trials.

[0260] In this study, a memory preservation model was used. The non-competitive NMDA receptor antagonist MK-801 (dizocilpine) was used to induce discrimination memory deficits. The efficacy of sGC stimulators in preventing MK-801-induced memory impairment was evaluated. The reference compound galantamine 1 mg / kg (intraperitoneal) significantly reversed discrimination deficits induced by MK-801 0.1 mg / kg (intraperitoneal), demonstrating the effectiveness of the test.

[0261] [, Materials and Methods , ] [animal] [] Adult male Langevin rats (275–299 g upon arrival from Envigo, Indianapolis, IN) were used in this study. Rats were housed in experimental rooms and assigned unique identification numbers (tail tags). Two rats were housed in polycarbonate cages with filtered tops and acclimatized for at least 7 days prior to testing. Animal rooms were maintained at a 12 / 12 h light / dark cycle (lights on at 07:00 EST), 22 ± 1°C, and approximately 50% relative humidity. Food and water were provided freely. All animals were examined, handled, and weighed prior to the study to ensure adequate health and minimize test-related nonspecific stress. Animals were randomly assigned to the entire treatment group. Experiments were conducted during the light cycle phases of the animals.

[0262] [Test compounds and reagents] [] The following compounds and reagents were used in these studies:

[0263] Dissolve MK-801 (0.1 mg / kg Sigma-Aldrich) in physiological saline and administer via intraperitoneal injection 15 minutes prior to NOR training. The dose volume is 1 ml / kg.

[0264] Galantamine (1 mg / kg; Tocris) was dissolved in saline and injected intraperitoneally 15 minutes before training. The dose volume was 1 ml / kg.

[0265] Will [Compound] [I]-

[20] (0.03, 0.3 and 1 mg / kg) were mixed in a medium (0.5% (w / w) methylcellulose and 0.5% (w / w) Tween 80 / ultrapure water) and administered orally at a dose of 2 ml / kg 60 min prior to NOR training.

[0266] The following groups were tested, with N=16 in each group (one rat was removed from the compound I-20 - 1 mg / kg - MK-801 group before the start of testing due to health issues): 1) Carrier-saline; 2) Carrier-MK-801 0.1 mg / kg; 3) Galantamine 1 mg / kg - MK-801; 4) Compound I-20, 0.03 mg / kg - MK-801; 5) Compound I-20, 0.3 mg / kg - MK-801; and 6) Compound I-20, 1 mg / kg - MK-801

[0267] [Compound] [I]-

[14] The compound solution and the compound mediator were prepared as Na+ salts in MilliQ water at concentrations of 0.01, 0.1, and 1 mg / kg and stored as frozen aliquots. The compound solution and the compound mediator were stored at -80°C and thawed fresh on each test day. The compound and mediator were administered orally 60 minutes before NOR training. The dose volume was 10 ml / kg.

[0268] Test the following groups [Compound] [I]-

[14] [,]In each group N=16: 1) physiological saline - MK-801 0.1 mg / kg; 2) galantamine 1 mg / kg - MK-801; 3) mediator - physiological saline; 4) mediator - MK-801; 5) compound I-14, 0.01 mg / kg - MK-801; 6) compound I-14, 0.1 mg / kg - MK-801; and 7) compound I-14, 1 mg / kg - MK-801

[0269] [Experimental Procedure] [] NOR tests were conducted in an open area (40 × 40 cm) in a quiet, dimly lit room. Each rat was tested individually, and olfactory / gustatory cues were removed by cleaning the testing area and test objects with 70% alcohol between the test and the rat. All training and testing trials were video-recorded and scored by observers unaware of the treatment.

[0270] On days 1 and 2, rats were allowed a 5-minute acclimatization period to freely explore the enclosure (without objects). On day 3 (training and testing day), rats were administered a mediator, saline, and / or compound solution according to the corresponding pretreatment, defined as the time between injection and the start of NOR training. Each animal was placed in a test enclosure containing two identical objects. Rats were placed in the same position within the enclosure, facing the same direction, and the time spent actively exploring the objects during the 3-minute training phase (T1) was recorded. After training, the rats were returned to their cages. The NOR test (T2) was performed 1 hour after T1. Each rat was returned to the test enclosure containing one familiar object and one novel object for 5 minutes, and the time spent exploring both objects was recorded. The order and position of the objects (left / right) presented in T2 were randomized among the rats to prevent bias relative to order or position preferences.

[0271] [Organization Collection] [] Approximately 10 minutes after T2 (135 minutes after drug administration), collect trunk blood into microcentrifuge tubes containing K2EDTA. Keep the blood tubes on ice for short-term storage. Centrifuge the tubes at 10,000 RPM for 10 minutes in a refrigerated centrifuge within 15 minutes. Extract the plasma and store the sample at -80°C until shipment to the client.

[0272] [Statistical Analysis] [] The data for the NOR test (T2) is represented as the identification index, which is defined as the ratio of the time spent exploring novel objects to the total time spent exploring both objects during the testing phase (novelty / (familiarity + novelty) × 100%).

[0273] for [Compound] [I]-

[14] The data were analyzed separately in two batches. The first batch included the saline-MK-801 group and the galantamine-MK-801 group. The data were analyzed using a t-test to assess the validity of the test. The second batch included the "sGC stimulator group", which included five treatment groups containing compound mediators (MilliQ water): mediator-saline, mediator-MK-801, compound I-14 0.01 mg / kg-MK-801, compound I-14 0.1 mg / kg-MK-801 and compound I-14 1 mg / kg-MK-801. The data of the second batch were analyzed by one-way ANOVA, followed by Fisher LSD post-hoc analysis in the time ranges of 0 to 1, 0 to 3 and 0 to 5 minutes. The significance was set at P<0.05. Nineteen animals with identification indices above 90% or below 30% were excluded due to a strong (non-memory) bias between the two objects. Two rats with a total exploration time of less than 10 seconds between the two objects were also excluded due to unreliable results (this is our NOR test criterion). Based on feedback from plasma analysis, one rat was removed due to suspected drug exposure. Subsequently, statistical outliers above or below the mean by two standard deviations were removed from further analysis. Based on these criteria, 1 to 6 rats were excluded from each experimental group (initially N=16) and from statistical analysis across all time ranges (0 to 1, 0 to 3, and 0 to 5 minutes).

[0274] for [Compound] [I]-

[20] [,] Data were analyzed using a one-way ANOVA followed by a Fisher LSD post-hoc analysis at time ranges of 0 to 1, 0 to 3, and 0 to 5 minutes, with significance set at P < 0.05. Animals with a discrimination index higher than 90% or lower than 30% were excluded due to strong (non-memory) bias between the two objects. Subsequently, statistical outliers above or below the mean by two standard deviations were removed from further analysis. Based on these criteria, 2 to 4 rats were excluded from each experimental group (initially N=16) and from all statistical analyses at all time ranges (0 to 1, 0 to 3, and 0 to 5 minutes).

[0275] [, result , ] [, , ] [a)] [Compound] [I-14] None of the rats in this study exhibited significant side effects at any dose. The rats maintained normal levels of alertness, activity, and object exploration.

[0276] Within the 0-1 minute time range, the t-test showed a significant difference between the saline-MK-801 group and the galantamine-MK-801 group (P<0.01), thus indicating the validity of this analysis. ANOVA of the compound groups showed a significant primary therapeutic effect on the identification index [F(4,56)=4.698, P<0.01]. Post-hoc analysis by Fisher LSD showed that within this time range, the median-saline group and the 1 mg / kg group... [Compound] [I]-

[14] Both groups in the MK-801 group showed significant differences from the mediator-MK-801 group (P<0.05 and P<0.01, respectively), indicating that MK-801 0.1 mg / kg induced significant memory loss and that compound I-14 1 mg / kg reversed the defect.

[0277] Within the 0-3 minute time range, the t-test showed a significant difference between the saline-MK-801 group and the galantamine-MK-801 group (P<0.001), indicating the validity of this analysis. ANOVA on the compound groups showed a significant primary therapeutic effect on the discrimination index [F(4,56)=5.113, P<0.01]. Post-hoc analysis showed a significant difference between the mordant-saline group and the mordant-MK-801 group within this time range (P<0.01), indicating that MK-801 0.1 mg / kg induced significant memory loss. Within this time range, 1 mg / kg... [Compound] [I]-

[14] Shows a trend toward reversing MK-801-induced amnesia (P<0.10).

[0278] Within the 0-5 minute time range, the t-test showed a significant difference between the saline-MK-801 group and the galantamine-MK-801 group (P<0.001), thus indicating the validity of this analysis. ANOVA of the compound groups showed a significant primary therapeutic effect on the identification index [F(4,56)=2.847, P<0.05]. Post-hoc comparative analysis showed that within this time range, 1 mg / kg... [Compound] [I]-

[14] It has a significantly higher identification index compared to the MK-801 group (P<0.05).

[0279] [b)] [Compound] [I-20] None of the rats in this study exhibited significant side effects at any dose. The rats maintained normal levels of alertness, activity, and object exploration.

[0280] ANOVA demonstrated a significant primary therapeutic effect on the identification index within the 0 to 1 min time range [F(5,77)=3.379, P<0.01]. Post-hoc analysis showed no significant difference between the mediator-MK-801 0.1 mg / kg group and the mediator-saline group within this time range (P>0.05). [Compound] [I]-

[20] The 1 mg / kg MK-801 group and the galantamine MK-801 group showed significantly higher identification indices than the carboxyl MK-801 group (P<0.01). Generally, data in the 0 to 1 min time range are relatively unstable; treatment in the 0 to 3 min and 0 to 5 min time ranges provides more reliable results.

[0281] Within the 0 to 3 min time range, ANOVA revealed a significant primary therapeutic effect on the recognition index [F(5,77)=3.922, P<0.01]. Post-hoc analysis showed that MK-801 0.1 mg / kg induced significant memory loss within this time range (carrier-saline group vs. carrier-MK-801 group, P<0.05). [Compound] [I]-

[20] The 1 mg / kg MK-801 group and the galantamine MK-801 group significantly reversed MK-801-induced memory deficits (Ps<0.001). [Compound] [I]-

[20] The 0.3 mg / kg MK-801 group also showed a trend of reversing MK-801-induced memory deficits within this time frame (P<0.10).

[0282] Within the 0-5 min time range, ANOVA demonstrated a significant primary therapeutic effect [F(5,77)=5.219, P<0.001]. Post-hoc analysis showed that MK-801 0.1 mg / kg induced significant memory loss, with the identification index approaching the chance level (50%). Galantamine (1 mg / kg), [Compound] [I]-

[20] 0.3 mg / kg and 1 g / kg significantly reversed MK-801-induced memory deficits (Ps<0.05, P<0.01 and Ps<0.001 compared with the MK-801 group). [Compound] [I] The [-20]0.03 mg / kg - MK-801 group also showed a trend of reversing MK-801-induced memory deficits within this time frame (P<0.10).

[0283] [] [Example]

[12] [:right] [Drowsiness-Awakening Drug in Rats with Implanted Telemetry Device] [EEG] [of] [Evaluate] [, , ] This study was conducted at PsychoGenics, Inc. The procedure was approved by the Laboratory Animal Management and Use Committee in accordance with the National Institutes of Health's guidelines for the management and use of laboratory animals.

[0284] [, Target , ] [, , ] This study was designed to evaluate [Compound] [I]-

[14] Specific drug EEG (electroencephalography) characteristics during sleep in young adult male SD rats with wireless implantation.

[0285] [, Materials and Methods , ] [, , ] [animal] [] Young, adult male Spög-Dolly (SD) rats (approximately 275–325 grams upon arrival) from Envigo (Indianapolis, IN) were used in this study. Upon arrival, each rat was assigned a unique identification number and housed in polycarbonate cages with micro-isolation filter tops at a rate of three rats per cage. All rats were examined and weighed before the start of the study to ensure adequate health and fitness. A 12 / 12 light / dark cycle was maintained throughout the study. Room temperature was maintained between 20°C and 23°C, with relative humidity maintained at approximately 50%. Food and water were provided freely during the study. After surgery, the rats were housed individually. After the recovery phase (7–10 days), the animals were transferred to the EEG recording room and placed on a DSI receiver for recording.

[0286] [Research Group] [] a. Agent A i. Pathway: PO ii. Volume: 10 ml / kg iii. Ingredient: Water b. 3 mg / kg [Compound] [I-14] i. Pathway: PO ii. Volume: 10 ml / kg iii. Preparation: 3 mg / kg in water c. 10 mg / kg [Compound] [I-14] i. Pathway: PO ii. Route: 10 ml / kg iii. Preparation: 10 mg / kg in water d. 30 mg / kg [Compound] [I-14] i. Pathway: PO ii. Route: 10 ml / kg iii. Preparation: 30 mg / kg in water

[0287] [] [Surgical Procedure] [] In short, the animal had a DSI telemetry device (F50-EET) implanted in three channel (lead pair) structures. Note the placement of the positive and negative leads at each location. Each of the EEG channels on the DSI transmitter acts as a differential input, which is used to measure the difference between the positive and negative leads. Forehead / parietal bone (right): anterior (+) 2 mm, lateral 2 mm; posterior (-) 4 mm, lateral 2 mm; Forehead / forehead (bilateral hemispherical): anterior (+) 2 mm, lateral 2 mm (right); anterior (-) 2 mm, lateral 2 mm (left); and neck EMG.

[0288] In summary, animals were anesthetized with 4% to 5% isoflurane and 1 L / min oxygen before surgery for induction, and maintained with a 1% to 2.5% isoflurane / 1 L / min oxygen mixture during surgery. The depth of anesthesia was monitored using hind limb withdrawal reflexes and respiratory rate measurements. The isoflurane concentration was continuously adjusted to maintain the surgical anesthesia level throughout the procedure. Ophthalmic lubricant was applied to the eyes using sterile swabs. The surgical site was prepared for sterile surgery by shaving, rinsing with chlorhexidine, and cleaning the skin three times with alcohol swabs. The abdomen, neck, and head were shaved and sterilized with chlorhexidine rinsing and three alternating applications of alcohol (2% chlorhexidine gluconate and 4% isopropanol). The anesthetized animal was then placed on a warm water circulating heating pad for surgical implantation of a telemetry device.

[0289] [Telemetry Implantation] A 3-4 cm incision is made along the midline of the top of the skull, extending approximately 1 cm posteriorly to the midpoint between the eyes and the base of the skull, extending into the midway dorsal neck region. Dissection is performed using a blunt instrument, creating a subcutaneous pouch from the posterior end of the incision by pushing alongside the connective tissue into the right ventral region. The pouch is flushed with sterile saline, and the transmitter is placed within it, with the lead extending out of the neck incision. A lead connected to the transmitter is measured, cut to size, and inserted into one of the dorsal neck muscles, secured in place with 5-0 silk sutures. This procedure is repeated for the second lead to ensure both leads are positioned in a straight line along the same muscle bundle, spaced 2-4 mm apart, and providing electromyographic (EMG) signals. The periosteum is then removed to expose skull landmarks, the anterior fontanelle, and the lambda suture tip. If necessary, the skull area is wiped with sterile saline and then dried with gauze or a sterile cotton swab (Q-tip). Use stereotactic coordinates of the regions of interest (posterior and parietal cortex) to mark the EEG leads. For each region of interest, drill one to two holes in the skull to ensure exposure of the dura mater. Insert dura mater contact screws into the holes and wrap the EEG leads around them (up to four holes). These screws must be in contact with the dura mater to detect EEG brain signals. Penetration through the dura mater will not cause adverse events and may improve the EEG signal (more brain contact with metal). Then, permanently seal the screws and surrounding skull surface area using the FLOW-It ALC compound. After the adhesive has fully dried, close the skin covering the skull with staples.

[0290] [Test Plan] [] This study followed a Monday-Thursday dosing / recording schedule, with baseline EEG recordings for 2 hours followed by the rats receiving a first dose of 0.3, 3, or 10 mg / kg between 7:50 and 8:00 AM. [Compound] [I]-

[14] Or mediator A (all orally) and a second dose of the same substance was administered approximately 12 hours later, followed by EEG for 12 hours. ● Start EEG recording at 6 AM (when the lights are on). ● Administer the medication at 7:50 AM (1 hour and 50 minutes after the lights are turned on). ● The second dose (of the same compound) was administered at 7:50 PM (1 hour and 50 minutes after lights out). ● Continue EEG recording until 7:50 AM the next day (24 hours after the first dose and 12 hours after the second dose).

[0291] Data were recorded using the Data Science International (DSI) data acquisition platform from rats moving freely in their cages. Recordings were taken during light and dark cycles. Light was maintained at 6 AM and 6 PM throughout the study. Animals were acclimatized to medication (carrier administration) prior to data collection.

[0292] Animals were tested in a crossover design with a clearance period of at least 72 hours between doses. On each test day, animals received the compound or mediator orally exactly 2 hours after the lights were turned on (6:00 AM) (7:50 AM). Data were recorded starting 2 hours before administration and continuing for 24 hours after administration.

[0293] [EEG] [Signal Evaluation] [] Ensure that the EEG does not contain line noise (50 or 60 Hz) or any continuous non-physiological frequency patterns that could be considered noise from an external power source. All EEG recordings are within the normal operating range (i.e., the EEG signal is within the normal operating amplitude range, typically greater than 100 µV and less than 500 µV, with no signal fidelity loss, usually achieved by reducing the observation by <100 µV).

[0294] [Sleep Score] [] The raw EEG recordings were manually scored using neuroscoping software (International Data Science) to identify sleep stages: active wakefulness, quiet wakefulness, NREM, and REM. The data were removed offline using a neurological scoring (DSI) method, and sleep stages were artificially assigned every 10 seconds using EEG, EMG, and spontaneous activity (LMA) using conventional methods as previously described (Ivarsson et al., 2005; Parmentier-Batteur et al., 2012; Leiser et al., 2014, 2015) using anterior parietal EEG, LMA, and EMG: active wakefulness (irregular low-amplitude EEG, and high EMG and LMA activity); quiet wakefulness (irregular low-amplitude EEG, and low EMG and no LMA activity); NREM (consisting of high-amplitude irregular waves with predominant δ (1 to 4 Hz), low EMG and no LMA); REM sleep (consisting of stable low-amplitude waves dominated by θ (4 to 8 Hz), almost no EMG and no LMA).

[0295] Sleep stage data were exported from a neurological scoring report template measuring sleep duration every 15 minutes (2 hours before administration to 4 hours after administration). The first sleep episode and the first REM episode were also directly reported from the template. Latency was calculated as the time interval between the first REM episode following the NREM episode (i.e., TREM - TNREM = REM latency). Hypnograms were generated using the percentage of time spent in each sleep stage across hourly time intervals. The time spent in each sleep state was calculated as the percentage of total time spent in that sleep state (mean ± standard error of the mean, SEM). Individual animal data were grouped by treatment group, sleep state, and frequency and exported to GraphPad PRISM for statistical analysis and visualization.

[0296] [Spectrum Analysis] [] Spectral analysis was performed using Matlab. Time-domain signals collected from multiple channels were fed into the DSI / Neurological Score database, and the EDF files were subsequently transferred to Matlab. Excel files labeled with baseline and specific post-dose timestamps were also transferred to Matlab for time locking. In Matlab, the Power Spectral Density (PSD) was calculated using the Welch method. Then, raw and relative spectral power were calculated for each of the six frequency bands (δ, θ, α, β, low γ, and high γ) and each 1 Hz subband. Data recorded 2 hours prior to compound administration were combined and defined as the "baseline." The percentage change relative to baseline was calculated based on each channel, individual, dose level, spectral band, and time segment. The mean raw, relative, and percentage changes for each frequency band were calculated for each group. Spectral analysis included quantifying the raw spectral power, relative spectral power, and percentage change in spectral power for each recorded EEG band (δ - 0.5 to 3.9 Hz, θ - 4 to 7.9 Hz, α - 8 to 11.9 Hz, β - 12 to 29.9 Hz, low γ - 30 to 49.9 Hz, and high γ - 50 to 100 Hz) for each rat. Additionally, the EEG spectrum from 1 to 100 Hz was represented as a line graph. These spectral data were provided separately to the user, and not all data is included in this document due to the large number of graphs. For clarity, the percentage change in each band over the time period following drug administration is presented.

[0297] [, result , ] [, , ] [Sleep] [] [Compound] [I]-

[14] Sleep patterns showed a significant reduction in REM and NREM in the high-dose (30 mg / kg) treatment group compared to the mediator (A) group. At the highest dose, an increase in quiet arousal was observed in the compound group. In this treatment group, the onset of NREM, REM, and REM latency was delayed. [Compound] [I]-

[14] The most profound effects occur at high doses, with the effects on quiet wakefulness (increased), REM (decreased), and NREM (decreased) lasting for several hours after administration.

[0298] [] [Spectrum Analysis] [] In QW, [Compound] [I]-

[14] (3 mg / kg) increased low γ from 0 to 180 minutes after administration, while at a dose of 30 mg / kg, it increased low γ from 0 to 240 minutes after administration and increased high γ from 0 to 180 minutes after administration. At 30 mg / kg, in NREM, the compound decreased δ, θ, α and increased low and high γ from approximately 0 to 300 minutes after administration.

[0299] [] [Example]

[13] [:] [Evaluate the compound] [I-14] [Long-term low-dose therapy in patients with Parkinson's disease and cognitive impairment] [MPTP] [Cognitive Role in a Diseased Macaque Model] This study is a non-GLP study in a macaque model of Parkinson's disease induced by long-term low-dose MPTP. This model is described in the literature (https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC3282499 / ). The study was designed to evaluate the effect of compound I-14 as a monotherapy on performance on the following cognitive tasks: variable delayed response (VDR), sustained performance (CPT), visual discrimination reversal learning (SDR), and object retrieval (OR). The study was conducted in animals previously treated with low-dose MPTP and known to have difficulty performing on the tasks mentioned above. These studies utilized four male cynomolgus macaques with MPTP-induced cognitive deficits.

[0300] This study consisted of four parts. Part 1 involved collecting baseline data from four animals on the four tasks under investigation. Part 2 involved administering a mediator (sterile filtered distilled water) for 16 days, with mediator data collected weekly for two weeks on each of the four tasks. Part 3 involved oral administration of compound I-14 daily for five days, followed by weekly data collection on each of the four tasks for two weeks, with compound I-14 (3.0 mg / kg) administered daily two hours before each test. The final part of the study included a nine-day clearance period, with weekly data collection on each of the four tasks for two weeks thereafter. After five days of pre-administration and two weeks of daily administration, a single dose of compound I-14 was evaluated as a monotherapy (3.0 mg / kg). The effects of treatment on cognitive performance were assessed two hours after daily administration and during the clearance period.

[0301] MPTP-HCl is administered intravenously at doses ranging from 0.05 mg / kg to 0.30 mg / kg, 2 to 3 times per week for several months. MPTP administration continues until cognitive deficits are observed, accompanied by minimal / mild Parkinson's disease motor impairment. An animal is considered "cognitively impaired" if it exhibits at least a 15% reduction in cognitive task performance relative to its pre-MPTP baseline. Because the response to MPTP is somewhat specific, MPTP is administered to achieve its effect (i.e., cognitive deficits followed by motor deficits), rather than for a specific duration of exposure or a specific cumulative dose.

[0302] SDR stands for Test of Cognitive Flexibility. In this task, three stimuli are presented simultaneously on a screen, one of which is arbitrarily designated as a positive (reward) stimulus, which produces a positive hue and reward upon contact, while a negative (non-reward) stimulus produces a different hue and a blank screen. The position of the stimuli on the screen varies pseudo-randomly across trials. Up to 300 trials are presented in each phase. The measurements recorded for each phase are: 1) the total number of trials required to learn the initial discrimination (i.e., achieving the specified 14 / 16 correct criterion) and 2) the total number of trials required to learn (i.e., the same criterion mentioned above) the reversal, where the previous negative (non-reward) stimulus is now a positive (reward) stimulus (i.e., achieving the criterion). The number of trials required to achieve the criterion for learning the reversal of discrimination is considered a measure of cognitive flexibility.

[0303] Compound I-14 monotherapy significantly improved efficacy in the SDR paradigm. CY3018 significantly reduced the number of trials required to achieve the criteria for learning discrimination reversal, which is designed to improve at least one cognitive state in these animals. Simple discrimination efficacy was not significantly altered by the administration of the causative agent or compound I-14. However, administration of compound I-14 significantly improved discrimination reversal learning efficacy (Figure 6).

[0304] Furthermore, during baseline, causative, and clearance tests, some animals failed to learn to recognize reversal; however, when tested with compound I-14, none of the animals failed to learn reversal, indicating that the compound has a positive effect on all tested animals.

Claims

1. Use of a compound of formula I or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating CNS diseases, health conditions or ailments in an individual in need: , wherein: The JC series is selected from the group consisting of hydrogen, halogens, and C1-6 alkyl; X is N or C (JC1); the JC1 series is selected from the group consisting of hydrogen, halogens, and C1-6 alkyl; each JB is independently selected from the group consisting of hydrogen, halogens, and C1-6 alkyl; the JD series is selected from the group consisting of hydrogen, halogens, and C1-6 alkyl; and n is an integer selected from 0, 1, 2, 3, or 4.

2. As claimed in claim 1, wherein the compound is represented by formula IA: , or a pharmaceutically acceptable salt thereof.

3. As requested in item 1, where JC1 is H, F, or Cl.

4. As requested in item 1, where JC1 is H.

5. As requested in item 1, where JC1 is F.

6. As claimed in claim 1, wherein the compound is represented by formula IB: , or a pharmaceutically acceptable salt thereof.

7. As requested in item 1, where n is 2 or 3.

8. As requested in item 1, where n is 0 or 1.

9. As claimed in claim 1, wherein each JB is independently H, F or C1-4 alkyl.

10. As claimed in claim 9, wherein each JB is independently F or methyl.

11. As requested in claim 7, wherein n is 2 and JB are both F or one of JB is F and the other is methyl.

12. As requested in claim 7, where n is 3 and two of JB are F and the other is methyl.

13. As requested in item 8, where n is 1 and JB is F.

14. As requested in item 8, where n is 0.

15. As requested in item 1, where JD is hydrogen.

16. As used in claim 1, wherein JD is F, Cl or methyl.

17. As requested in item 1, where JD is F.

18. As requested in item 1, where JC is H or F.

19. As requested in item 1, where JC is H.

20. For any of the uses of claims 1 to 19, wherein the CNS disease is Alzheimer's disease.

21. As requested in claim 20, wherein the Alzheimer's disease is mild to moderate or moderate to severe.

22. For any of the uses of claims 1 to 19, wherein the CNS disease is cognitive impairment.

23. For any of the uses of claims 1 to 19, wherein the CNS disease is dementia.

24. For the purposes of any of claims 1 to 19, wherein the CNS disease is subjective cognitive impairment (SCI).

25. For any of the uses of claims 1 to 19, wherein the CNS disease is cognitive aging.

26. For any of the uses of claims 1 to 19, wherein the CNS disease is vascular dementia.

27. For any of the uses of claims 1 to 19, wherein the CNS disease is mixed dementia.

28. For any of the uses of claims 1 to 19, wherein the CNS disease is Parkinson's disease.

29. For the purposes of any of claims 1 to 19, wherein the CNS disease is mild cognitive impairment.

30. For any of the claims 1 to 19, wherein the CNS disease is a traumatic (closed or open) head puncture injury, traumatic brain injury (TBI), non-traumatic stroke, aneurysm, hypoxia, or other brain injury.

31. For any of the uses of claims 1 to 19, wherein the CNS disease is stroke.

32. As requested in claim 31, wherein the CNS disease is ischemic stroke.

33. The use of any of claims 1 to 19, wherein the drug further comprises additional therapeutic agents.

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