Selecting treatment of patients with FADS-1 mediated diseases or disorders with FADS-1 inhibitors

Abstract

The present disclosure provides techniques for acquiring FADS1 activity in a patient. Also provided are techniques for determining the suitability of treating a patient with a FADS1 regulatory (e.g., inhibitory) compound. The determination may be made by analyzing one or more biological indicators of a FADS1-mediated disease or disorder in the subject. The one or more biological indicators may include one or more of a ratio of polyunsaturated fatty acids ("PUFAs") in the subject, a relative abundance of one or more cell types, a relative abundance of one or more differentially expressed genes ("DEG") (or gene characteristics, e.g., RNA for such DEG), and / or a relative abundance of one or more metabolites. Also disclosed are methods of using FADS1 inhibitors in methods of treating metabolic disorders and obesity.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 384,862, filed on November 23, 2022, and U.S. Provisional Patent Application No. 63 / 592,804, filed on October 24, 2023. Each of the foregoing applications is incorporated herein by reference in its entirety for all purposes. Technical Field

[0003] The present disclosure provides techniques for determining the level of fatty acid desaturase 1 ("FADS1") activity in a subject using one or more biomarkers. Also disclosed are methods of treating FADS1-mediated metabolic disorders and obesity in a subject in need of treatment using a FADS1 inhibitor. In several embodiments, the subject in need of treatment is a subject having one or more biomarkers of a FADS1-mediated disease or FADS1-mediated disorder. In several embodiments, the subject in need of treatment is one having increased FADS1 activity relative to a healthy population. Background Art

[0004] Polyunsaturated fatty acids ("PUFAs") play important physiological functions in the human body. PUFAs serve as energy sources and structural components of cell membranes. PUFAs also regulate genes and are biosynthetic precursors for other physiologically relevant biomolecules, such as eicosanoids and endocannabinoids. Eicosanoids are signaling molecules that have multiple functions and, in particular, regulate the human inflammatory response. Endocannabinoids (e.g., N-arachidonylethanolamine (AN) and 2-arachidonylglycerol (2-AG) are endogenous ligands for cannabinoid receptors and have been identified as playing a role in food intake and energy balance.

[0005] Figure 1(d) shows the relevant portion of the metabolic pathway for a particular PUFA, linoleic acid ("LA"), which leads, inter alia, to the formation of anti-inflammatory and pro-inflammatory eicosanoids and endocannabinoids. The desaturases that catalyze certain steps in the conversion of LA to dihomo-gamma-linolenic acid ("DGLA") and arachidonic acid ("AA") are delta-6-desaturase ("D6D," encoded by the gene fatty acid desaturase 2 ("FADS2")) and delta-5-desaturase ("D5D," encoded by the gene fatty acid desaturase 1 ("FADS1")). Selective inhibition of D5D activity reduces the amount of AA produced while increasing the amount of DGLA. This pharmacological intervention reduces the downstream production of, for example, pro-inflammatory eicosanoids and endocannabinoids and leads to the accumulation of anti-inflammatory eicosanoids, both of which can generally improve conditions associated with inflammation and can improve energy balance. This is particularly relevant in subjects with high LA intake (e.g., those exposed to a Western diet).

[0006] The FADS1-3 locus has been associated with numerous metabolic traits in human genome-wide association studies, including fasting glucose, plasma lipids, and body weight. In addition to human genetic evidence supporting a role for FADS1 / D5D in metabolic disorders, FADS1 knockout ("KO") mice exhibit a phenotype that is resistant to diet-induced obesity, including low body fat content, improved glycemic control, and reduced circulating lipid levels. Furthermore, FADS1 KO mice are resistant to the development of atherosclerotic plaques.

[0007] Desaturase enzyme activity has been linked to a variety of diseases, particularly metabolic and cardiovascular diseases such as obesity, diabetes, nonalcoholic steatohepatitis ("NASH"), dyslipidemia, and coronary artery disease. Therefore, pharmacological inhibition of D5D is an interesting target for the treatment of metabolic, cardiovascular, and other diseases. Summary of the Invention

[0008] One aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information from the subject comparing the level of a biological indicator for a FADS1-mediated disease with a reference level of the biological indicator; wherein the biological indicator is the AA to DGLA ratio; and if the level of the biological indicator in the subject is above the reference level of the biological indicator, administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound. Another aspect of the present disclosure provides a method for detecting a FADS1-mediated disease or disorder in a subject, the method comprising: receiving information from the subject comparing the level of a biological indicator for a FADS1-mediated disease with a reference level of the biological indicator; wherein the biological indicator is the AA to DGLA ratio. Another aspect of the present disclosure provides a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1-mediated disease or disorder, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is the AA to DGLA ratio; and if the level of the biological indicator in the subject is above the reference level of the biological indicator, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound.

[0009] Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 5:1. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 6:1. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 7:1. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 15:2. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 8:1. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 17:2. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the subject has an AA to DGLA ratio of equal to or at least about 9:1.

[0010] Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a PUFA; and if the comparison of the level of the biological indicator in the subject to the reference level of the biological indicator determines that the subject is a subject who would benefit from treatment with a FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound. Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: if the level of the biological indicator for the FADS1-mediated disease in the subject is above the reference level of the biological indicator, administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound; wherein the biological indicator is a measured level of a PUFA. Another aspect of the present disclosure provides a method for detecting a FADS1-mediated disease or disorder in a subject, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a PUFA. Another aspect of the present disclosure provides a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1-mediated disease or disorder, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a PUFA; and if the subject is determined by comparing the level of the biological indicator in the subject to the reference level of the biological indicator to be a subject that would benefit from treatment with the FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound. In some embodiments, the PUFA is linoleic acid, arachidonic acid, gamma-linoleic acid, adrenic acid, dihomo-gamma-linolenic acid, docosapentaenoic acid n-6, alpha-linolenic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosahexaenoic acid, or a combination of the foregoing.

[0011] Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a metabolite; and if the subject is determined by comparing the level of the biological indicator in the subject to the reference level of the biological indicator to be a subject that would benefit from treatment with a FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound. Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: if the level of the biological indicator for the FADS1-mediated disease in the subject is above the reference level of the biological indicator, administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound; wherein the biological indicator is a measured level of a metabolite. Another aspect of the present disclosure provides a method for detecting a FADS1-mediated disease or disorder in a subject, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a metabolite. Another aspect of the present disclosure provides a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1-mediated disease or disorder, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a metabolite; and if the subject is determined by comparing the level of the biological indicator in the subject to the reference level of the biological indicator to be a subject who would benefit from treatment with the FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound.In some embodiments, the metabolite is cholesterol, free cholesterol, total cholesterol, cholesterol ester C20:4, malate, α-ketoglutarate, mannose, glucose, erythro-dihydrosphingosine (d18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadiene, 4-hydroxydihydrosphingosine (t18:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18:1, C24:0), ceramide (d18:1, C24:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadiene, 4-hydroxydihydrosphingosine (t18:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18:1, C24:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadiene, ... :0), thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxyheptadecatrienoic acid (C17: [5,8,10]3), 14,15-dihydroxyeicosatrienoic acid (C20: cis [5,8,11]3), 11-hydroxyeicosatetraenoic acid (C20: cis [5,8,12,14]4), 13-hydroxyoctadecadienoic acid (13-HODE) (C18: cis [9] trans

[11] 2), arachidonic acid (C20: cis [5,8,11,14]4), docosahexaenoic acid (C22: cis [4,7,10,13,16,19]6), dihomo-γ-isothiazolinone (DISODIUM HYDROXYPROPYL HYDROLYZED ... Linoleic acid (C20: cis[8,11,14]3), gamma-linolenic acid (C18: cis[6,9,12]3), docosapentaenoic acid (C22: cis[7,10,13,16,19]5), eicosapentaenoic acid (C20: cis[5,8,11,14,17]5), docosatetraenoic acid (C22: cis[7,10,13,16]4), stearic acid (C18:0), tryptophan, kynurenic acid, xanthurenic acid, histidine, leucine, isoleucine, valine, 3-hydroxyisobutyric acid, glutamic acid, threonine, cysteine, sarcosine, plasma triglycerides, taurochenodeoxycholic acid, taurocholic acid, lysophosphatidylcholine (C18:0 ), lysophosphatidylcholine (C20:4), lysophosphatidylcholine (C17:0), lysophosphatidylethanolamine (C22:5), phosphatidylcholine (C18:0, C22:6), phosphatidylcholine (C18:0, C20:3), phosphatidylcholine (C18:1, C18:2), phosphatidylcholine (C16:1, C18:2), phosphatidylcholine (C18:0, C18:2), phosphatidylcholine (C16:0, C20:5), phosphatidylcholine (C16:0, C16:0), glycerol-3 phosphate, choline plasmalogen (C18, C20:4), inositol, inositol phospholipids, glycerol phosphate, phospholipid fraction, or a combination of the foregoing.

[0012] Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a differentially expressed gene ("DEG"); and if the subject is determined by comparing the level of the biological indicator for the subject to the reference level of the biological indicator to be a subject who would benefit from treatment with a FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound. Another aspect of the present disclosure provides a method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising administering to the subject a therapeutically effective amount of a FADS1 inhibitor compound if the level of the biological indicator for the FADS1-mediated disease in the subject is above the reference level of the biological indicator; wherein the biological indicator is a measured level of a DEG. Another aspect of the present disclosure provides a method for detecting a FADS1-mediated disease or disorder in a subject, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a DEG. Another aspect of the present disclosure provides a FADS1 inhibitor compound for use in a method of treating a subject in need of treatment for a FADS1-mediated disease or disorder, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a DEG; and if the subject is determined by comparing the level of the biological indicator in the subject to the reference level of the biological indicator to be a subject that would benefit from treatment with the FADS1 inhibitor compound, administering to the subject a therapeutically effective amount of the FADS1 inhibitor compound. In some embodiments, the DEGs are Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Akr1d1, Aldh1a1, Mmp19, Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel1l3, Agap2, 4833411C07Rik, Elov4, Fat 3. Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36 827, Man2a2, Nudt18, Plagl1, TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b,Tagln2, Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, Cyp2b9 Hao2, Cyp2b13, Cyp2a22, Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, Fitm1, Lp ar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, Ly6f, Foxi1, Cryg c, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, Ppp2r5b, Slc6a7, Gpr50 Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Gm18537, Pclo, Adrb3, Gm38394, A C154232.2, Cadps, Adgrb2, Gm45470, Sdr9c7, Dsg1c, Slc17a1, Ces1c, Gss 1810008I18Rik, Tlcd1, Snrk, Akr1c20, Gm19950, Ttr, Cbfa2t3, Acat2, P mvk, Abcd3, Acacb, Arhgap27, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl 19. Tcf7, Xkrx, Trim46, Zfp369, Zfp871, Pcdhb21, Gm14288, Uprt, Atm chs2, Cped1, Gm38357, Cck, Ckap2, Gm4419, 1600015I10Rik, Sez6I2, Prnd Gm16702, S100a8, Pcdh12, Malat1, Kcnq1ot1, ArI4c, Gm42549, Gm37310. Gm37776, Atp2a1, Ckm, Tnnt3, Adipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9, Ce bpa, Dgat1, Dgat2, Elovl3, Fas, Scd1, Srebf1, Hilpda, Lipe, Mgll, Plin1 Plin4, Pnpla2, Pnpla3, Ldah, Cs, Gckr, Me1, Pck1, Pdk4, Irs1, Acaa1a, A cads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, C.S yp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, and Tnf.

[0013] Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a cell type; and if the comparison of the level of the biological indicator in the subject to the reference level of the biological indicator determines that the subject is a subject who would benefit from treatment with a FADS1 inhibitor compound, administering a therapeutically effective amount of the FADS1 inhibitor compound to the subject. Another aspect of the present disclosure provides a method for treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: if the level of the biological indicator for the FADS1-mediated disease in the subject is above the reference level of the biological indicator, administering a therapeutically effective amount of a FADS1 inhibitor compound to the subject; wherein the biological indicator is a measured level of a cell type. Another aspect of the present disclosure provides a method for detecting a FADS1-mediated disease or disorder in a subject, the method comprising: receiving information comparing the level of a biological indicator for a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a cell type. Another aspect of the present disclosure provides a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1-mediated disease or disorder, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject with a reference level of the biological indicator; wherein the biological indicator is a measured level of a cell type; and if the subject is determined by comparing the level of the biological indicator in the subject with the reference level of the biological indicator to be a subject that would benefit from treatment with the FADS1 inhibitor compound, administering a therapeutically effective amount of the FADS1 inhibitor compound to the subject. In several embodiments, the cell type is an adipocyte ("Adipo"), a B cell ("Bcell"), an endothelial cell ("Endo"), a hepatocyte ("Hep"), a Kupffer cell ("Kupff"), a bone marrow cell ("Myel"), a natural killer cell ("NK"), a T cell ("Tcell"), or a combination of the foregoing.

[0014] As disclosed elsewhere herein, several embodiments relate to treating FADS1-mediated diseases in subjects in need of treatment. Several embodiments disclosed herein relate to selecting patients for treatment using FADS1 inhibition (e.g., by administering a FADS1 inhibitor). In several embodiments, in this selection step, subjects who are more likely to respond favorably to treatment are selected for treatment. In several embodiments, patients with a higher likelihood of successful treatment are then treated using, for example, a FADS1 modulating compound (e.g., a FADS1 inhibitor compound). In several embodiments, selection is based on patient data collected prior to administration of the FADS1 inhibitor. In several embodiments, the patient data is associated with measurements of one or more biological indicators of FADS1 activity (e.g., increased FADS1 activity). In several embodiments, selection is based on patient data collected before, concurrently with, and / or after administration of the FADS1 inhibitor. In several embodiments, FADS1-mediated diseases or disorders can be treated by inhibiting the FADS1 enzyme using one or more methods disclosed herein.

[0015] Several embodiments disclosed herein provide methods for identifying a subject with increased FADS1 activity. In several embodiments, the method comprises measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject. In several embodiments, the one or more biological indicators comprise one or more of the following: the proportion of PUFAs in the subject, the relative abundance of one or more cell types, the relative abundance of one or more DEGs (or gene signatures, such as RNA for such DEGs), and / or the relative abundance of one or more metabolites. In several embodiments, the method further comprises administering a FADS1 inhibitor compound to the subject.

[0016] Several embodiments disclosed herein provide a method for identifying a subject in need of treatment with a FADS1 inhibitor compound. In several embodiments, the method comprises measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject. In several embodiments, the one or more biological indicators comprise, consist of, or consist essentially of one or more of the following: the proportion of PUFAs in the subject, the relative abundance of one or more cell types, the relative abundance of one or more DEGs or gene signatures, and / or the relative abundance of one or more metabolites. In several embodiments, the method further comprises administering a FADS1 inhibitor compound to the subject.

[0017] Several embodiments disclosed herein provide methods for reducing weight, reducing body mass index, treating obesity, treating metabolic disorders, treating cardiovascular disorders, treating diabetes, treating dyslipidemia, and / or treating non-alcoholic steatohepatitis ("NASH") in a subject. In several embodiments, the method comprises measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject. In several embodiments, the one or more biological indicators comprise, consist of, or consist essentially of one or more of the proportion of PUFAs in the subject, the relative abundance of one or more cell types, the relative abundance of one or more DEGs or gene signatures, and / or the relative abundance of one or more metabolites. In several embodiments, the method comprises administering a FADS1 inhibitor compound to the subject.

[0018] In several embodiments, as disclosed herein, the methods disclosed herein further comprise administering a FADS1 inhibitor compound to a subject. In several embodiments, the methods disclosed herein further comprise administering a dose of labeled DGLA to a subject and then measuring the ratio of labeled AA to labeled DGLA. In several embodiments, the ratio of labeled AA to labeled DGLA allows calculation of the inhibitory concentration achieved with the FADS1 inhibitor. In several embodiments, the inhibitory concentration achieved with the FADS1 inhibitor is used to calculate the inhibitor dosage appropriate for the subject.

[0019] Another aspect of the present disclosure provides a method of measuring the ratio of AA to DGLA in a subject; wherein the AA is isotopically labeled and the DGLA is isotopically labeled; wherein the ratio of AA to DGLA is measured by administering a dose of labeled DGLA to the subject and then measuring the ratio of labeled AA to labeled DGLA.

[0020] Another aspect of the present disclosure provides a compound represented by the following structure:

[0021]

[0022] Where: Each "*" symbol indicates that the isotope can be enriched 13 C position; and at least one "*" position is isotopically enriched 13 C.

[0023] Another aspect of the present disclosure provides a compound represented by the following structure:

[0024]

[0025] Where: Each "*" symbol indicates that the isotope can be enriched 13 C position; and at least one "*" position is isotopically enriched 13C.

[0026] Another aspect of the present disclosure provides a compound represented by the following structure:

[0027]

[0028] Where: X is a halogen; each "*" symbol indicates that it can be isotopically enriched 13 C position; and at least one "*" position is isotopically enriched 13 C.

[0029] Another aspect of the present disclosure provides a method of making a compound represented by the following structure:

[0030]

[0031] The method comprises making the following compound

[0032]

[0033] Reacts with KC*N; where each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13 C; and wherein X is a suitable leaving group. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figures 1(a)-(c) provide data demonstrating that FADS1 activity is increased in humans and rodent models of obesity. Figure 1(a) provides a flow diagram showing the n-3 PUFA and n-6 PUFA pathways. As shown, FADS1 mediates the conversion of n-6 PUFA (DGLA to AA) and n-3 PUFA (eicosatetraenoic acid (ETA) to eicosapentaenoic acid (EPA)). Figure 1(b) provides a comparison of the plasma AA / DGLA ratio in lean, overweight, and obese human men, as a surrogate for FADS1 activity; n = 8-41 / group; data are presented as minimum to maximum values. Figure 1(c) compares the AA / DGLA ratio in lean and high-fat diet (HFD)-induced obese (DIO) male C57Bl / 6 mice following 12 weeks of HFD, as a surrogate for FADS1 activity. n = 8 / group. Figures 1(b,c) t-test. *P < 0.05, ****P < 0.00001, respectively.

[0035] Figure 1(d) is a scheme showing the metabolic pathway of a certain PUFA (LA), which leads to the formation of, inter alia, anti-inflammatory and pro-inflammatory eicosanoids and endocannabinoids.

[0036] Figures 2(a)-(h) provide data supporting that Fads1 KO mice are resistant to HFD-induced obesity and have improved metabolic profiles. Figure 2(a) shows a comparison of age-matched WT and Fads1 KO mice on a 12-week HFD. The data demonstrate that Fads1 KO mice are resistant to HFD-induced weight gain. Figure 2(b) shows that Fads1 KO mice have lower fat mass, and Figure 2(c) shows that Fads1 KO mice have lower lean body mass relative to their WT littermates. Figure 2(d) shows that food intake measured at 7 weeks in Fads1 KO mice was not significantly altered relative to WT mice. Figure 2(e) shows that Fads1 KO mice have lower insulin levels relative to WT mice. Figure 2(f) shows that Fads1 KO mice have improved glucose tolerance during a GTT relative to WT mice. Figure 2(g) shows that Fads1 KO mice have lower cholesterol relative to WT mice. Figure 2(h) shows that no differences were found in plasma triglyceride levels; n = 8-9 per group. Data are expressed as mean ± SEM. (a-c) Two-way ANOVA with Sidak's multiple comparison test; (b-h) t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001.

[0037] Figures 3(a)-(l) provide data supporting that HFD-induced obese Fads1 KO mice have a lower respiratory exchange ratio (RER) and increased dark cycle energy expenditure. Fads1 KO and WT mice were placed on an HFD for 12 weeks; n = 24 mice / group. Figures 3(a,b) show oxygen consumption for the two groups, Figures 3(c,d) show carbon dioxide production for the two groups, Figures 3(e,f) show RER measurements for the two groups, and Figures 3(g,h) show activity measurements for the two groups at 0, 6, and 12 weeks of HFD feeding. In Figures 3(a,c,e,g), each data point represents a rolling average of six time points, with the dark cycle (6:00 PM to 6:00 AM) indicated by the outlined box. In Figures 3(b,d,f,h), measurements were averaged and shown as mean ± SEM for each day or night over time. Two-way ANOVA with Sidak's multiple comparison test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001. Figure 3 (i1) provides a graph of energy expenditure (kcal / hour) during the light and dark cycles compared to body weight after 6 and 12 weeks of HFD feeding; least squares multiple linear regression model including body weight and genotype.

[0038] Figure 4(a)-(d) shows RNA-seq analysis of liver, epididymal ("EPI") white adipose tissue ("WAT"), and inguinal ("ING") WAT of Fads1 KO mice compared to WT when fed a HFD for 12 weeks. Figure 4(a) provides a volcano plot representing DEGs in Fads1 KO compared to WT in liver, EPI WAT, and ING WAT; genes with a Benjamini-Hochberg (BH) adjusted P value <0.01 and a fold change ≥2 or ≤0.5 are marked with color. Figure 4(b) shows an analysis of the average cellular composition of liver, EPI WAT, and ING WAT of Fads1 KO and WT mice by SCDC using scRNA-Seq of the corresponding mouse organs as a reference. Cell type abbreviations are as follows: adipocytes ("Adipo"), B cells ("Bcell"), endothelial cells ("Endo"), hepatocytes ("Hep"), Kupffer cells ("Kupff"), myeloid cells ("Myel"), natural killer cells ("NK"), T cells ("Tcell"). Kyoto Encyclopedia of Genes and Genomes ("KEGG") differentially altered metabolic pathways analysis in liver ( FIG. 4( c) ) and EPI WAT ( FIG. 4( d) ) of Fads1 KO mice relative to WT mice is provided; the number of unique genes with significantly altered expression in each pathway is shown; n = 4-8 / group.

[0039] Figures 5(a)-(b) demonstrate that Fads1 knockout mice fed a high-fat diet have altered metabolic gene expression, increased adiponectin levels, and reduced inflammatory markers in adipose tissue. Figure 5(a) provides a comparison of metabolic gene expression changes in EPI WAT, ING WAT, and liver of Fads1 KO mice relative to WT littermates; genes with a Benjamini-Hochberg-adjusted P value < 0.01 and a fold change ≥ 2 or ≤ 0.5 are color-coded. Figure 5(b) provides a comparison of adiponectin, leptin, PAI-1, and MCP-1 levels in plasma, EPI WAT, and ING WAT in WT versus Fads1 KO mice; (a) Data are shown as minimum to maximum values. n = 3-8 / group. t-test *P < 0.05, **P < 0.01, ***P < 0.001.

[0040] Figures 6(a)-(b) demonstrate altered fatty acid composition in plasma lipid subfractions of Fads1 KO mice. Figure 6(a) provides data on fatty acid concentrations in total plasma and each of the four major lipid subfractions (phospholipids, free fatty acids, cholesterol esters, and triglycerides) from HFD-fed WT and Fads1 KO mice. Figure 6(b) provides fatty acid composition in total plasma and each of the four lipid subfractions from WT and Fads1 KO mouse plasma. (a-b) n = 3 samples / genotype (each sample pooled from 2 mice). (a) Data are shown as minimum to maximum values. *P < 0.05, **P < 0.01, ***P < 0.001, t-test.

[0041] Figures 7(a)-(c) show metabolomic analysis of Fads1 KO mice on a HFD relative to WT mice. Figure 7(a) provides a comparison of plasma metabolite profiles in Fads1 KO mice compared to WT mice after 11 weeks of HFD feeding. Ratios of Fads1 KO metabolite levels relative to WT are shown. Only metabolites with ratios ≥1.15 or ≤0.85 and P values ≤0.05 are shown; n = 4-6 / group. t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.00001. Figure 7(b) provides a heat map of the top-ranked diseases and affected biological functions in the IPA based on the metabolic profiles of Fads1 KO mice on a HFD relative to WT mice. Figure 7(c) provides a heat map comparing common and differentially altered diseases and biological functions in the plasma metabolome and transcriptome of liver, EPI WAT, and ING WAT in Fads1 KO mice relative to WT mice. In Figure 7(bc), only diseases and biological functions with P values < 0.05 are shown.

[0042] Figures 8(a)-(i) present data demonstrating that a small molecule FADS1 inhibitor reduced body weight and improved metabolic parameters in DIO mice. Compound A in Figure 8(a) is a FADS1 inhibitor, and as shown in Figure 8(b), it exhibits >1200X selectivity for human FADS1 over human FADS2, as measured by FADS1 and FADS2 cell-based assays. Similar efficacy was observed in the mouse FADS1 cell-based assay in Figure 8(b) and in Figure 8(c), which provides an in vivo FADS1 efficacy assay in mice. DIO mice treated daily with 10 mg / kg and 30 mg / kg of Compound A for 54 days resulted in reduced body weight, as shown in Figure 8(d), and decreased residual hepatic FADS1 activity in vivo, as shown in Figure 8(e), without affecting food intake, as shown in Figure 8(f). Compound A treatment reduced insulin (Figure 8(g)), cholesterol (Figure 8(h)), and triglyceride levels (Figure 8(i)) (n = 7-10 per group). Data are presented as mean ± SEM. Figure 8(d) Two-way ANOVA with Sidak's multiple comparison test; Figure 8(ei) One-way ANOVA with Dunnett's multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001. ANOVA, analysis of variance; DIO, diet-induced obesity; FADS1, fatty acid desaturase 1; FADS2, fatty acid desaturase 2; MW, molecular weight; POC, percentage of control; SEM, standard error of the mean.

[0043] Figure 9 (a)-(j) provides data of indirect calorimetry of DIO mice treated with compound A or mice treated with vehicle. Figure 9 (a) provides a weight comparison, Figure 9 (b) provides the relationship between light cycle and dark cycle energy consumption compared to body weight, Figure 9 (c, d) provides oxygen consumption, Figure 9 (e, f) provides carbon dioxide production, Figure 9 (g, h) provides RER, and Figure 9 (i, j) provides activity. DIO mice were treated with vehicle or compound A (30 mg / kg) for 42 days and measured at the specified time. In Figure 9 (c, e, g, i), each data point represents a rolling mean of 3 days, and the dark cycle (6:00 pm to 6:00 am) is shown by an outline box. In Figure 9 (d, f, h, j), the measured values are averaged and displayed as the mean ± SEM of each day or night over time. Figure 9 (b) is a least squares multiple linear regression model including body weight and treatment. Figure 9 (a, d, f, h, j) Two-way ANOVA with Sidak's multiple comparison test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001. n = 10-11 mice / group.

[0044] Figure 10 (a)-(d) provides RNA-seq analysis of age-matched Fads1KO and WT DIO mice treated with vehicle or 30 mg / kg Compound A for 54 days. Figure 10 (a) provides a radar chart of commonly altered genes in liver, EPI WAT, and ING WAT of Fads1KO mice in Compound A-treated mice relative to WT vehicle-treated mice. Figure 10 (b) provides the average cell composition in liver, EPI, and ING WAT of Fads1KO, WT+Compound A, and WT mice by SCDC analysis using scRNA-Seq of the corresponding mouse organs as a reference. Cell types are abbreviated as follows: Adipo, Bcell, Endo, Hep, Kupff, Myel, NK, Tcell. Ingenuity Pathway Analysis ("IPA") analysis identified common and unique pathways affected by Fads1 KO or inhibition in Figure 10 (c) EPI WAT and Figure 10 (d) liver; n = 6-9 / group.

[0045] Figures 11(a)-(c) present data demonstrating that no differences in FADS2 activity were observed in obese humans and mice. Higher FADS1 activity, but not higher FADS2 activity, was observed in HFD-induced obese mice. Figure 11(a) shows that no differences in the plasma GLA / LA ratio (a surrogate marker of FADS2 n-6 activity) were observed in overweight or obese human males relative to lean subjects; n = 8-41 / group. Figure 11(b) similarly shows that no differences in the plasma GLA / LA ratio were observed in DIO mice fed an HFD for 12 weeks (n = 7-8 / group). In Figure 11(c), SC-26196, a FADS2-specific inhibitor, did not affect FADS1 activity as demonstrated by in vitro assays of human FADS1, FADS2, and mouse FADS1 activity. Data for Figures 11(a,b) are presented as minimum to maximum values; t-test. Figures 11(c,d) n = 4-8 / group; data are presented as mean ± SEM.

[0046] Figure 12 It was shown that inhibition of FADS2 activity by SC-26196 affects not only the plasma GLA / LA ratio but also the plasma AA / DGLA ratio in DIO mice. Figure 12 One-way ANOVA with Dunnett's multiple comparison test, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0047] Figures 13(a)-(h) show that DIO mice treated with Compound A exhibit altered metabolic gene expression, increased adiponectin levels, and decreased inflammatory markers. Figure 13(a) liver, Figure 13(b) EPI WAT, and Figure 13(c) ING WAT provide a comparison of metabolic gene expression changes in WT DIO mice treated with vehicle (WT), WT DIO mice treated with Compound A (30 mg / kg) (WT+A), and Fads1 KO DIO mice treated with vehicle (KO). n = 6-9 / group. Figure 13(d) liver and 13(e) EPI WAT provide IPA analysis of the top-ranked predicted upstream regulators in WT DIO mice treated with Compound A (WT+A) and Fads1 KO DIO mice (KO) relative to WT DIO mice treated with vehicle (WT); predicted upstream regulators and activation z scores with P overlap values <0.01 are indicated by shading. Figure 13(f) provides a comparison of adiponectin, leptin, PAI-1, and MCP-1 levels in plasma, epiwastewater (EPIWAT), and ingwastewater (INGWAT) in WT, Compound A-treated WT, and Fads1 KO DIO mice; n = 7-9 per group. Data for Figures 13(ac,f) are presented from minimum to maximum. One-way ANOVA with Dunnett's multiple comparison test compared to vehicle-treated WT. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.00001. Figure 13 (g) provides a volcano plot representing DEGs in liver, EPI, and ING WAT of diet-induced obese mice in Fads1 KO mice compared to WT and 30 mg / kg Compound A-treated (WT+A) groups compared to vehicle-treated WT (WT); the number of genes with a Benjamini-Hochberg-adjusted P value <0.01 and a fold change ≥2 or ≤0.5 is color-coded and provided in a Venn diagram format representing the number of genes uniquely or similarly altered in liver and adipose tissue in Fads1 KO versus WT or WT+A versus WT cohorts. Figure 13 (h) provides a comparison of liver PAI-1 and MCP-1 levels in WT, Compound A-treated WT, and Fads1 KO DIO mice.

[0048] Figures 14(a) and (b) provide data showing that FADS1 inhibition reduces endocannabinoid levels in liver and adipose tissue. Comparison of endocannabinoids, 2-arachidonoyl-glycerol ("2-AG"), and anandamide ("AEA") levels in liver (Figure 14(a)) and ING WAT (Figure 14(b)) of vehicle-treated WT DIO mice (WT), Compound A (30 mg / kg)-treated WT DIO mice (WT+A), and vehicle-treated Fads1 KO DIO mice (KO); Data are presented as minimum to maximum values; n = 8-6 / group. One-way ANOVA with Dunnett's multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001.

[0049] Figures 15(a)-(f) show that FADS1 inhibition increases the expression of PPARα target genes without significant effects on hepatic triglyceride content and steatosis-related genes. A comparison of hepatic triglyceride levels in WT and Fads1 KO mice receiving a normal diet (Figure 15(a)) or a 12-week HFD (Figure 15(b)) is provided; n = 4-8 / group. A comparison of hepatic triglyceride levels in vehicle-treated Fads1 KO mice and WT DIO mice after 8 weeks of treatment with vehicle or 10 or 30 mg / kg Compound A is provided (Figure 15(c)); data are presented in the form of minimum to maximum values; n = 8 / group. Figure 15(d) provides liver histological assessments of WT DIO mice and age-matched vehicle-treated Fads1 KO DIO mice treated with vehicle or Compound A for 24 days. Compared to WT vehicle-fed DIO mice, WT DIO mice and Fads1 KO DIO mice treated with Compound A (10 mg / kg) had similar or smaller vacuoles (H&E staining) and reduced lipid content (Oil Red O staining), indicating that Compound A treatment did not worsen hepatic steatosis or loss of Fads1 expression; n = 3 / group. Representative images are shown. Hepatic gene expression of steatosis-related genes is shown in Figure 15(e), and PPARα target genes in Fads1 KO mice relative to WT littermates after 8 weeks of HFD are shown in Figure 15(f). Genes with a BH-adjusted P value < 0.01 and a fold change ≥ 2 or ≤ 0.5 are marked with color. n = 4-6 / group. Figure 15(a,b) t-test; Figure 15(c) one-way ANOVA with Dunnett's multiple comparison test.

[0050] Figure 16 (a)-(b) shows that the fatty acid composition in the plasma lipid subfraction of DIO mice treated with compound A changes relative to that of vehicle-treated DIO mice. Figure 16 (a) provides the fatty acid concentration of each of the total plasma and 4 major lipid subfractions (phospholipids, free fatty acids, cholesterol esters and triglycerides) of WT DIO mice treated with vehicle (WT) or 30 mg / kg compound A (WT+A) for 54 days. Figure 16 (b) provides the fatty acid composition of each of the total plasma and 4 plasma lipid subfractions of WT DIO mice (WT+A) treated with vehicle (WT) and 30 mg / kg compound A. (ab) n=3 samples / group (each sample is combined from 2 mice). (a) Data are shown as minimum to maximum values. T-test *P<0.05, **P<0.01, ***P<0.001.

[0051] Figure 17a provides the results for compounds BE. 13 C5-DGLA to 13 The % conversion data of C5-AA, such as using the 13 C5-DGLA and 13 C5-AA measurement. Figure 17b provides the % inhibition for compounds BE, as measured using plasma collected from 13 C5-DGLA and 13 C5-AA Measurements A dose response curve for Compound F was generated and is shown in Figure 17c. DETAILED DESCRIPTION

[0052] Chronic and persistent inflammation contributes to the pathology of obesity and its comorbidities. FADS1 is a key enzyme that synthesizes AA from DGLA. AA is a precursor to many pro-inflammatory eicosanoids. On the other hand, DGLA is a precursor to several anti-inflammatory eicosanoids. Several embodiments disclosed herein relate to the treatment of FADS1-mediated diseases, conditions, and / or disorders. In several embodiments, the method includes selecting a subject suffering from a FADS1-mediated disease or disorder. In several embodiments, the subject is selected for treatment based on the level of FADS1 activity in the subject. In several embodiments, the subject has elevated FADS1 activity. In several embodiments, when the subject has elevated FADS1 activity, the method includes administering a FADS1 inhibitor to the subject. In several embodiments, the subject's FADS1 activity is measured before, during, or after treatment. In several embodiments, the subject's FADS1 activity is indicated by the AA / DGLA ratio in the subject (or by other biological indicators disclosed elsewhere herein). In several embodiments, the level of FADS1 activity in a subject is compared to the level of FADS1 activity in a subject (or subject population) that does not have a FADS1-mediated disease or disorder. In several embodiments, FADS1 activity is elevated in obese subjects (e.g., humans), as indicated by the AA / DGLA ratio (or other biological indicators disclosed elsewhere herein). Inhibiting FADS1 activity may alleviate obesity and its metabolic comorbidities. U.S. Application Publication Nos. US2021 / 0171529 and US2021 / 0188874 describe a family of FADS1 inhibitor compounds as agents for treating metabolic or cardiovascular diseases.

[0053] The section headings used herein are for organizational purposes only and should not be considered as limiting the subject matter described. Features disclosed under one heading (e.g., composition) may be used in combination with features disclosed under different headings (methods of manufacture or treatment).

[0054] definition

[0055] The following definitions are provided to assist in understanding the scope of the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs.

[0056] As used herein, the term "pharmaceutically acceptable" means generally accepted for use in subjects, particularly humans.

[0057] As used herein, the term "pharmaceutically acceptable excipient" refers to various ingredients that can be combined with the compounds or salts disclosed herein to prepare pharmaceutical compositions or formulations. Typically, excipients include, but are not limited to, diluents, colorants, vehicles, anti-adherents, glidants, disintegrants, flavorings, coatings, binders, sweeteners, lubricants, adsorbents, preservatives, and the like.

[0058] The term "pharmaceutically acceptable salt" refers to a salt of a compound that has the desired pharmacological activity of the parent compound and is not undesirable for its final use in biology or other aspects. Pharmaceutically acceptable salts include, for example, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid) or with organic acids (e.g., acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid). Pharmaceutically acceptable salts also include, for example, salts formed when the acidic protons present in the parent compound are replaced by metal ions (e.g., alkali metal ions, alkaline earth metal ions, or aluminum ions) or when associated with organic bases (e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, dicyclohexylamine). Additionally, salts of the compounds described herein can exist in hydrated or anhydrous forms or as solvates with other solvent molecules.

[0059] As used herein, the term "patient" or "subject" refers to humans and mammals, including but not limited to primates, cows, sheep, goats, horses, dogs, cats, rabbits, rats and mice. In one embodiment, the subject is a human.

[0060] As used herein, the term "therapeutically effective amount" refers to that amount of a compound disclosed herein that will elicit the biological or medical response of a tissue, system, or subject that is being sought by the researcher, veterinarian, medical doctor, or other clinician.

[0061] As used herein, the term "relative abundance" is a comparison of the abundance of a particular feature in a test subject (e.g., a subject in need of treatment) relative to the abundance of the same feature in a non-test subject or a non-test subject population. The non-test subject or non-test subject population can include healthy subjects and / or those who do not need treatment.

[0062] As used herein, the term "healthy subject" is a person having the average characteristics of a healthy population of individuals. Healthy is given its simple and ordinary meaning and includes subjects who lack one or more or all FADS1-mediated diseases and / or disorders. A healthy population can be a population that includes individuals with a body mass index of less than about 25, about 22.5, or about 20. A healthy population can be a population that includes individuals with a body mass index greater than about 18.5. A healthy population can be a population that includes individuals who do not suffer from obesity, metabolic disorders, cardiovascular disorders, diabetes, dyslipidemia, and / or NASH.

[0063] The term "isotopologue" refers to a substance whose chemical structure differs from a specific compound of the present invention only in its isotopic composition.

[0064] Isotopically enriched compounds are disclosed herein. It will be appreciated that some variation in natural isotopic abundance occurs in labeled compounds synthesized as disclosed herein, depending on the sources of the chemical materials used in the synthesis. Therefore, preparations of labeled compounds will contain small amounts of labeled isotopologues. In the labeled compounds disclosed herein, any atom not specifically designated as a particular isotope is intended to represent any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated as "C" or "carbon" or is undesignated, that position is understood to have carbon at its naturally abundant isotopic composition. Furthermore, unless otherwise stated, when a position is specifically designated as " 13 C” or “isotopically enriched”, the position is understood to have an abundance of at least 13 The natural abundance of C is 100 times higher 13 C.

[0065] As used herein, the term "isotopic enrichment factor" refers to the ratio of the isotopic abundance to the natural abundance of a particular isotope. In several embodiments, the compounds of the present invention have a relative abundance of 1% for each of the specified isotopes. 13 The C atoms have an isotopic enrichment factor equal to or greater than about 100, 500, 1000, 1500, 2000, 2500, 3000, 4000, 5000, or a range including and / or spanning the above values. In several embodiments, the isotopically enriched compounds of the present invention have an isotopic enrichment factor equal to or greater than about 100, 500, 1000, 1500, 2000, 2500, 3000, 4000, 5000, or a range including and / or spanning the above values. 13 The C atoms have an isotopic enrichment factor equal to or greater than about 100, 500, 1000, 1500, 2000, 2500, 3000, 4000, 5000, or a range including and / or spanning the above values.

[0066] As used herein, the term "halogen" refers to -F, -Cl, -Br, or -I.

[0067] As known to those skilled in the art, certain compounds disclosed herein can exist in one or more tautomeric forms. Because a chemical structure can only be used to represent one tautomeric form, it is understood for convenience that reference to a compound having a given structural formula includes other tautomers having the structural formula.

[0068] When referring to numerical values, the term "or a range including and / or spanning the aforementioned values" (and variations thereof) is meant to include any range including or spanning the aforementioned values. To illustrate, when a value is expressed as "20, 30, 40, 50, or a range including and / or spanning the aforementioned values," this includes each specific value provided (e.g., 20, 30, 40, and / or 50) or any range spanning or including any two values provided (e.g., 20 to 50, 20 to 40, 20 to 30, 30 to 50, 30 to 40, or 40 to 50).

[0069] The following description provides context and examples, but should not be construed as limiting the scope of the invention as encompassed by the claims appended hereto or any other application claiming priority to this specification. No single component or combination of components is essential or indispensable. Any feature, structure, component, material, step, or method described and / or illustrated in any embodiment of this specification may be used with or in place of any feature, structure, component, material, step, or method described and / or illustrated in any other embodiment of this specification.

[0070] introduction

[0071] Dietary intake and endogenous synthesis of PUFAs, as well as their physiological regulation, influence human health and disease. The two major classes of PUFAs, ω-3 (n-3) and ω-6 (n-6) fatty acids ("FAs"), can be ingested or synthesized endogenously from primary precursors, n-3α-linolenic acid ("ALA") or n-6LA, which are FAs that mammals cannot synthesize. These FAs are sequentially metabolized by fatty acid desaturases and fatty acid elongases. The first rate-limiting enzyme is FADS2, also known as D6D, because it desaturates LA or ALA at the sixth carbon, converting them to gamma-linolenic acid ("GLA") or stearidonic acid, respectively (Figure 1(a)). Subsequent elongation of GLA and stearidonic acid produces DGLA and ETA, respectively (Figure 1(a)). DGLA and ETA are desaturated by FADS1 (also known as D5D), because FADS1 desaturates at the fifth carbon of the fatty acid chain; the corresponding products of FADS1 enzymatic action are n-6AA and n-3EPA acids (Figure 1(a)). Both AA and EPA are extensively metabolized by enzymes, including cyclooxygenase and lipoxygenase, to form biologically active eicosanoids, such as prostaglandins, thromboxanes, and leukotrienes. These eicosanoids play important roles in metabolism and inflammation, with AA-derived eicosanoids primarily having pro-inflammatory effects, while EPA-derived eicosanoids have anti-inflammatory properties.

[0072] Obesity and its associated comorbidities have become a global public health problem. The rise in obesity rates over the past few decades is partly due to a Westernized diet, which has a high ratio of n-6 to n-3 (approximately 10:1 to 20:1). This high ratio may contribute to the development of inflammation, cardiovascular disease, cancer, and autoimmune diseases. Excessive n-6 PUFAs (such as AA) lead to excessive amounts of proinflammatory eicosanoids and other oxylipins. Obesity and metabolic disorders may be partly the result of excessive eicosanoid-mediated inflammatory damage. Humans rely on the intake and production of PUFAs through FADS2 and FADS1 (encoded by their respective genes, FADS2 and FADS1). Differences in FADS1 expression between different alleles at the FADS locus have enabled the identification of numerous associations with traits and diseases from genome-wide association studies (GWAS). Notably, the FADS locus has been shown to have the highest association with AA concentration to date (P = 3 × 10 -971 ), more specifically, with AA / DGLA concentration (P = 2 × 10 -361). The minor C allele of the FADS1 single nucleotide polymorphisms ("SNPs") rs174556 and rs174547 was associated with reduced FADS1 activity and reduced body weight or waist circumference. Furthermore, improved metabolic phenotypes were observed in Fads1 KO mice when fed a normal diet or HFD. Taken together, these FADS1-related phenotypes in humans and mice suggest that reducing FADS1 activity may have therapeutic value in the treatment of obesity and related metabolic comorbidities.

[0073] In view of the foregoing, several embodiments provided herein relate to methods for determining whether a subject is a candidate for treatment with a FADS1 inhibitor compound. In several embodiments, a candidate for treatment with a FADS1 inhibitor compound may be a subject suffering from or at risk for a FADS1-mediated disorder or disease. In several embodiments, the method comprises measuring the level of a biological indicator in a biological sample from the subject. In several embodiments, the level of the biological indicator provides information regarding whether the patient suffers from or is at risk for a FADS1-mediated disease or disorder. In several embodiments, the level of the biological indicator is compared to a reference level of the biological indicator from a different subject (or subject population) who does not suffer from (or is not at risk for developing) a FADS1-mediated disease or condition. In several embodiments, if the level of the biological indicator in the subject indicates that the subject is a candidate for treatment with a FADS1 inhibitor compound (e.g., suffers from or is at risk for a FADS1-mediated condition), the FADS1 inhibitor compound is administered to the subject. In several embodiments, the biological indicator is one or more PUFAs (or a ratio of PUFAs). In several embodiments, the biological indicator is the relative abundance of one or more cell types in the subject. In several embodiments, the biological indicator is the relative abundance of one or more DEGs. In some embodiments, the biological indicator is the relative abundance of one or more metabolites. In some embodiments, the FADS1 inhibitor compound is a FADS1-inhibiting small molecule. In some embodiments, by inhibiting FADS1, the availability of AA and the production of pro-inflammatory eicosanoids are limited and / or reduced.

[0074] Methods of treatment and FADS1 inhibitor compounds for use in treatment

[0075] Example 1 provided herein is a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0076] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0077] Among them, the biological indicator is the ratio of arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA); and

[0078] If the level of the biological marker in the subject is above the reference level for the biological marker, a therapeutically effective amount of a FADS1 inhibitor compound is administered to the subject.

[0079] Example 2 provided herein is a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1 mediated disease or disorder, the method comprising:

[0080] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0081] Among them, the biological indicator is the ratio of arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA); and

[0082] If the level of the biological indicator in the subject is higher than the reference level of the biological indicator, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0083] Embodiment 3 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 5:1.

[0084] Embodiment 4 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 6:1.

[0085] Embodiment 5 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 7:1.

[0086] Embodiment 6 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 15:2.

[0087] Embodiment 7 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 8:1.

[0088] Embodiment 8 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 17:2.

[0089] Embodiment 9 provided herein is the method or compound of embodiment 1 or 2, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 9:1.

[0090] Example 10 provided herein is a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0091] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0092] wherein the biological marker is a measured level of linoleic acid, gamma-linoleic acid, adrenic acid, docosapentaenoic acid n-6, alpha-linolenic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof; and

[0093] If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0094] Example 11 provided herein is a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1 mediated disease or disorder, the method comprising:

[0095] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0096] wherein the biological marker is a measured level of linoleic acid, gamma-linoleic acid, adrenic acid, docosapentaenoic acid n-6, alpha-linolenic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof; and

[0097] If the subject is determined to be a subject that would benefit from treatment with the FADS1 inhibitor compound by comparing the level of the biological marker in the subject to the reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0098] Embodiment 12 provided herein is the method or compound of embodiment 10 or 11, wherein when the level of the biological indicator of the subject is higher than the reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

[0099] Embodiment 13 provided herein is the method or compound of embodiment 10 or 11, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is lower than the reference level of the biological indicator.

[0100] Example 14 provided herein is a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0101] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0102] The biological indicator is the following measurement levels: plasma cholesterol, free cholesterol, total cholesterol, cholesterol ester C20:4, malate, α-ketoglutarate, mannose, glucose, erythro-dihydrosphingosine (d18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadecadiene, 4-hydroxydihydrosphingosine (t18:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18:1 , C24:0), thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxyheptadecatrienoic acid (C17: [5,8,10]3), 14,15-dihydroxyeicosatrienoic acid (C20: cis [5,8,11]3), 11-hydroxyeicosatetraenoic acid (C20: cis [5,8,12,14]4), 13-hydroxyoctadecadienoic acid (13-HODE) (C18: cis [9] trans

[11] 2), arachidonic acid (C20: cis [5,8,11,14]4), docosahexaenoic acid (C22: cis [4,7,10,13,16,19]6), dihomo-gamma -linolenic acid (C20: cis[8,11,14]3), γ-linolenic acid (C18: cis[6,9,12]3), docosapentaenoic acid (C22: cis[7,10,13,16,19]5), eicosapentaenoic acid (C20: cis[5,8,11,14,17]5), docosatetraenoic acid (C22: cis[7,10,13,16]4), stearic acid (C18:0), tryptophan, kynurenic acid, xanthurenic acid, histidine, leucine, isoleucine, valine, 3-hydroxyisobutyric acid, glutamic acid, threonine, cysteine, sarcosine, plasma triglycerides, taurochenodeoxycholic acid, taurocholic acid, lysophosphatidylcholine (C18:0 ), lysophosphatidylcholine (C20:4), lysophosphatidylcholine (C17:0), lysophosphatidylethanolamine (C22:5), phosphatidylcholine (C18:0, C22:6), phosphatidylcholine (C18:0, C20:3), phosphatidylcholine (C18:1, C18:2), phosphatidylcholine (C16:1, C18:2), phosphatidylcholine (C18:0, C18:2), phosphatidylcholine (C16:0, C20:5), phosphatidylcholine (C16:0, C16:0), glycerol-3 phosphate, choline plasmalogen (C18, C20:4), inositol, inositol phospholipids, glycerol phosphate, phospholipid fraction, or a combination of the foregoing;

[0103] If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0104] Example 15 provided herein is a FADS1 inhibitor compound for use in a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0105] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0106] The biological indicator is the following measurement levels: plasma cholesterol, free cholesterol, total cholesterol, cholesterol ester C20:4, malate, α-ketoglutarate, mannose, glucose, erythro-dihydrosphingosine (d18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadecadiene, 4-hydroxydihydrosphingosine (t18:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18:1 , C24:0), thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxyheptadecatrienoic acid (C17: [5,8,10]3), 14,15-dihydroxyeicosatrienoic acid (C20: cis [5,8,11]3), 11-hydroxyeicosatetraenoic acid (C20: cis [5,8,12,14]4), 13-hydroxyoctadecadienoic acid (13-HODE) (C18: cis [9] trans

[11] 2), arachidonic acid (C20: cis [5,8,11,14]4), docosahexaenoic acid (C22: cis [4,7,10,13,16,19]6), dihomo-gamma -linolenic acid (C20: cis[8,11,14]3), γ-linolenic acid (C18: cis[6,9,12]3), docosapentaenoic acid (C22: cis[7,10,13,16,19]5), eicosapentaenoic acid (C20: cis[5,8,11,14,17]5), docosatetraenoic acid (C22: cis[7,10,13,16]4), stearic acid (C18:0), tryptophan, kynurenic acid, xanthurenic acid, histidine, leucine, isoleucine, valine, 3-hydroxyisobutyric acid, glutamic acid, threonine, cysteine, sarcosine, plasma triglycerides, taurochenodeoxycholic acid, taurocholic acid, lysophosphatidylcholine (C18:0 ), lysophosphatidylcholine (C20:4), lysophosphatidylcholine (C17:0), lysophosphatidylethanolamine (C22:5), phosphatidylcholine (C18:0, C22:6), phosphatidylcholine (C18:0, C20:3), phosphatidylcholine (C18:1, C18:2), phosphatidylcholine (C16:1, C18:2), phosphatidylcholine (C18:0, C18:2), phosphatidylcholine (C16:0, C20:5), phosphatidylcholine (C16:0, C16:0), glycerol-3 phosphate, choline plasmalogen (C18, C20:4), inositol, inositol phospholipids, glycerol phosphate, phospholipid fraction, or a combination of the foregoing;

[0107] If the subject is determined to be a subject that would benefit from treatment with the FADS1 inhibitor compound by comparing the level of the biological marker in the subject to the reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0108] Embodiment 16 provided herein is the method or compound of embodiment 14 or 15, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is higher than the reference level of the biological indicator.

[0109] Embodiment 17 provided herein is the method or compound of any one of embodiments 14 to 16, wherein the biological indicator is plasma cholesterol, free cholesterol, total cholesterol, cholesterol ester C20:4, mannose, glucose, erythro-dihydrosphingosine (d18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadiene, 4-hydroxydihydrosphingosine (t18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadiene, 4-hydroxysphingosine (t18:1), 8:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18:1, C24:0), thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxyheptadecatrienoic acid (C17: [5,8,10]3), 14,15-dihydroxyeicosatrienoic acid (C20: cis [5,8,11]3), 11-hydroxyeicosatetraenoic acid (C20: cis [5,8,12,14]4), 13-hydroxydeca Octadecadienoic acid (13-HODE) (C18: cis[9]trans

[11] 2), arachidonic acid (C20: cis[5,8,11,14]4), docosahexaenoic acid (C22: cis[4,7,10,13,16,19]6), docosapentaenoic acid (C22: cis[7,10,13,16,19]5), eicosapentaenoic acid (C20: cis[5,8,11,14,17]5), docosatetraenoic acid (C22: cis[7,10,13,16]4), stearic acid (C18: ), cysteine, plasma triglycerides, taurochenodeoxycholic acid, taurocholic acid, lysophosphatidylcholine (C18:0), lysophosphatidylcholine (C20:4), lysophosphatidylcholine (C17:0), phosphatidylcholine (C18:0, C22:6), phosphatidylcholine (C18:0, C20:3), phosphatidylcholine (C18:0, C18:2), phosphatidylcholine (C16:0, C20:5), choline plasmalogen (C18, C20:4), inositol, inositol phospholipids, phosphoglycerol, phospholipid fraction, or a combination of the foregoing.

[0110] Embodiment 18 provided herein is the method or compound of embodiment 14 or 15, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is lower than the reference level of the biological indicator.

[0111] Embodiment 19 provided herein is the method or compound of any one of embodiments 14 to 15 and 18, wherein the biological indicator is malate, alpha-ketoglutarate, dihomo-gamma-linolenic acid (C20: cis [8, 11, 14] 3), gamma-linolenic acid (C18: cis [6, 9, 12] 3), tryptophan, kynurenic acid, xanthurenic acid, histidine, leucine, isoleucine, valine, 3-hydroxyisobutyric acid, glutamic acid, threonine, sarcosine, lysophosphatidylethanolamine (C22:5), phosphatidylcholine (C18:1, C18:2), phosphatidylcholine (C16:1, C18:2), phosphatidylcholine (C16:0, C16:0), glycerol-3 phosphate, or a combination of the foregoing.

[0112] Example 20 provided herein is a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0113] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0114] The biological indicator is the measured level of differentially expressed genes (DEGs);

[0115] Among them, the DEGs are Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Akr1d1, Aldh1a1, Mmp19, Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel1l3, Agap2, 4833411C07Rik, Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Tagln2, Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, Cyp2b9, Hao2, Cyp2b13, Cyp2a22, Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, Fitm1, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, Ly6f, Foxi1, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, Ppp2r5b, Slc6a7, Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Gm18537, Pclo, Adrb3, Gm38394, AC154232.2. Cadps, Adgrb2, Gm45470, Sdr9c7, Dsg1c, Slc17a1, Ces1c, Gss, 1810008I18Rik, Tlcd1, Snrk, Akr1 c20, Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acacb, Arhgap27, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl19, Tcf7, Xkrx, Trim46, Zfp369, Zfp871, Pcdhb21, Gm14288, Uprt, Atm, Dchs2, Cped1, Gm3 8357, Cck, Ckap2, Gm4419, 1600015I10Rik, Sez6I2, Prnd, Gm16702, S100a8, Pcdh12, Malat1, Kcnq1o t1, ArI4c, Gm42549, Gm37310, Gm37776, Atp2a1, Ckm, Tnnt3, Adipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9 , Cebpa, Dgat1, Dgat2, Elovl3, Fas, Scd1, Srebf1, Hilpda, Lipe, Mgll, Plin1, Plin4, Pnpla2, Pnpla 3, Ldah, Cs, Gckr, Mel, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, Tnf, or a combination thereof; and

[0116] If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0117] Example 21 provided herein is a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1 mediated disease or disorder, the method comprising:

[0118] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0119] The biological indicator is the measured level of differentially expressed genes (DEGs);

[0120] Among them, the DEGs are Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Akr1d1, Aldh1a1, Mmp19, Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel1l3, Agap2, 4833411C07Rik, Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Tagln2, Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, Cyp2b9, Hao2, Cyp2b13, Cyp2a22, Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, Fitm1, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, Ly6f, Foxi1, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, Ppp2r5b, Slc6a7, Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Gm18537, Pclo, Adrb3, Gm38394, AC154232.2. Cadps, Adgrb2, Gm45470, Sdr9c7, Dsg1c, Slc17a1, Ces1c, Gss, 1810008I18Rik, Tlcd1, Snrk, Akr1 c20, Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acacb, Arhgap27, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl19, Tcf7, Xkrx, Trim46, Zfp369, Zfp871, Pcdhb21, Gm14288, Uprt, Atm, Dchs2, Cped1, Gm3 8357, Cck, Ckap2, Gm4419, 1600015I10Rik, Sez6I2, Prnd, Gm16702, S100a8, Pcdh12, Malat1, Kcnq1o t1, ArI4c, Gm42549, Gm37310, Gm37776, Atp2a1, Ckm, Tnnt3, Adipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9 , Cebpa, Dgat1, Dgat2, Elovl3, Fas, Scd1, Srebf1, Hilpda, Lipe, Mgll, Plin1, Plin4, Pnpla2, Pnpla 3, Ldah, Cs, Gckr, Mel, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, Tnf, or a combination thereof; and

[0121] If the subject is determined to be a subject that would benefit from treatment with the FADS1 inhibitor compound by comparing the level of the biological marker in the subject to the reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0122] Embodiment 22 provided herein is the method or compound of embodiment 20 or 21, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is higher than the reference level of the biological indicator.

[0123] Embodiment 23 provided herein is the method or compound of any one of embodiments 20 to 22, wherein the DEG is Serpinbl a, Gna14, Serpina3m, Hsd11bl, Cyp2c29, Hsd11bl, Akrld1, Aldh1al, or a combination of the foregoing.

[0124] Embodiment 24 provided herein is the method or compound of any one of embodiments 20 to 23, wherein the DEG is Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, or a combination of the foregoing.

[0125] Embodiment 25 provided herein is the method or compound of any one of embodiments 20 to 24, wherein the DEG is TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Tagln2, or a combination of the foregoing.

[0126] Embodiment 26 provided herein is the method or compound of any one of embodiments 20 to 25, wherein the DEG is Cyp2b9, Fadsl, Hao2, Cyp2bl3, Cyp2a22, Acnat2, Ildr2, Rpl10a-psl, Tm6sf2, or a combination of the foregoing.

[0127] Embodiment 27 provided herein is the method or compound of any one of embodiments 20 to 26, wherein the DEG is Ly6f, Foxil, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, or a combination of the foregoing.

[0128] Embodiment 28 provided herein is the method or compound of any one of embodiments 20 to 27, wherein the DEG is Slc6a7, Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Mfap2, or a combination of the foregoing.

[0129] Embodiment 29 provided herein is the method or compound of any one of embodiments 20 to 28, wherein the DEG is Sdr9c7, Dsg1c, Slc17a1, Acnat2, Ces1c, Gss, Hsd11b1, or a combination of the foregoing.

[0130] Embodiment 30 provided herein is the method or compound of any one of embodiments 20 to 29, wherein the DEG is Adam7, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl19, Tcf7, Xkrx, Trim46, or a combination of the foregoing.

[0131] Embodiment 31 provided herein is the method or compound of any one of embodiments 20 to 30, wherein the DEG is Cck, Ckap2, Gm4419, 1600015I10Rik, Sez6I2, Prnd, Gm16702, S100a8, or a combination of the foregoing.

[0132] Embodiment 32 provided herein is the method or compound of embodiment 20 or 21, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is lower than the reference level of the biological indicator.

[0133] Embodiment 33 provided herein is the method or compound of any one of embodiments 20 to 21 and 32, wherein the DEG is Tmem86a, Elovl2, Agap2, Chkb, Tnfaip8l1, H2afj, Sel113, 4833411C07Rik, or a combination of the foregoing.

[0134] Embodiment 34 provided herein is the method or compound of any one of embodiments 20-21 and 32-33, wherein the DEG is Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, or a combination of the foregoing.

[0135] Embodiment 35 provided herein is the method or compound of any one of embodiments 20 to 21 and 32 to 34, wherein the DEG is Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, or a combination of the foregoing.

[0136] Embodiment 36 provided herein is the method or compound of any one of embodiments 20-21 and 32-34, wherein the DEG is Fitm1, Tmem86a, Agap2, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, or a combination of the foregoing.

[0137] Embodiment 37 provided herein is the method or compound of any one of embodiments 20-21 and 32-34, wherein the DEG is Ppp2r5b.

[0138] Embodiment 38 provided herein is the method or compound of any one of embodiments 20 to 21 and 32 to 34, wherein the DEG is Gm18537, Aacs, Pclo, Adrb3, Gm38394, AC154232.2, Cadps, Adgrb2, Gm45470, or a combination of the foregoing.

[0139] Embodiment 39 provided herein is the method or compound of any one of embodiments 20-21 and 32-34, wherein the DEG is 1810008I18Rik, Tlcd1, Snrk, Akrlc20, Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acss2, Acacb, Arhgap27, or a combination of the foregoing.

[0140] Embodiment 40 provided herein is the method or compound of any one of embodiments 20 to 21 and 32 to 34, wherein the DEG is Zfp369, Zfp871, Pcdhb21, Adrb3, Gm14288, Uprt, Atm, Dchs2, Cped1, Gm38357, or a combination of the foregoing.

[0141] Embodiment 41 provided herein is the method or compound of any one of embodiments 20 to 21 and 32 to 34, wherein the DEG is Pcdh12, Malat1, Kcnq1ot1, ArI4c, Gm38394, Gm42549, AC154232.2, Gm37310, Gm37776, Atp2a1, Ckm, Tnnt3, or a combination of the foregoing.

[0142] Embodiment 42 provided herein is the method or compound of any one of embodiments 20 to 41, wherein the DEG is Mmp19, Gyp27a1, Tymp, or a combination of the foregoing.

[0143] Embodiment 43 provided herein is the method or compound of any one of embodiments 20 to 42, wherein the DEG is Adipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9, Cebpa, Dgat1, Dgat2, Elov13, Fas, Scd1, Srebf1, Adrb3, Hilpda, Lipe, Mg11, Plin1, Plin4, Pnpla2, Pnpla3, Ld ah, Cs, Gckr, Mel, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, Tm6sf2, Tnf, or a combination of the foregoing.

[0144] Embodiment 44 provided herein is a method or compound of any one of embodiments 20 to 43, wherein the DEG is a combination of the DEG disclosed in Figure 4a, Figure 5a, Figure 10a, Figure 13a, Figure 13g, Figure 15e, or Figure 15f, or the DEG disclosed above. For example, in several embodiments, the DEG is the DEG disclosed in Figure 4a. In several embodiments, the DEG is the DEG disclosed in Figure 5a. In several embodiments, the DEG is the DEG disclosed in Figure 10a. In several embodiments, the DEG is the DEG disclosed in Figure 13a. In several embodiments, the DEG is the DEG disclosed in Figure 13g. In several embodiments, the DEG is the DEG disclosed in Figure 15e. In several embodiments, the DEG is the DEG disclosed in Figure 15f. In several embodiments, the DEG is a combination of the DEG disclosed above or DEG. In several embodiments, the DEG is the DEG disclosed in any other figure provided herein.

[0145] Example 45 provided herein is a method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising:

[0146] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0147] wherein the biological indicator is a measured level of one or more cell types in the subject, wherein the cell type is adipocytes (Adipo), B cells (Bcell), endothelial cells (Endo), hepatocytes (Hep), Kupffer cells (Kupff), bone marrow cells (Myel), natural killer cells (NK), T cells (Tcell), or a combination thereof; and

[0148] If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0149] Example 46 provided herein is a FADS1 inhibitor compound for use in a method of treating a subject in need thereof having a FADS1 mediated disease or disorder, the method comprising:

[0150] receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator;

[0151] wherein the biological indicator is a measured level of one or more cell types in the subject, wherein the cell type is adipocytes (Adipo), B cells (Bcell), endothelial cells (Endo), hepatocytes (Hep), Kupffer cells (Kupff), bone marrow cells (Myel), natural killer cells (NK), T cells (Tcell), or a combination thereof; and

[0152] If the subject is determined to be a subject that would benefit from treatment with the FADS1 inhibitor compound by comparing the level of the biological marker in the subject to the reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject.

[0153] Embodiment 47 provided herein is the method or compound of embodiment 45 or 46, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is above the reference level of the biological indicator.

[0154] Embodiment 48 provided herein is the method or compound of any one of embodiments 45 to 46 and 47, wherein the cell type is Adipo, Myel, or a combination thereof.

[0155] Embodiment 49 provided herein is the method or compound of embodiment 45 or 46, wherein the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound when the level of the biological indicator in the subject is lower than the reference level of the biological indicator.

[0156] Embodiment 50 provided herein is the method or compound of any one of embodiments 45 to 46 and 49, wherein the cell type is a B cell, Endo, NK, T cell, or a combination thereof.

[0157] Embodiment 51 provided herein is a method or compound of any one of embodiments 1 to 50, wherein the FADS1-mediated disease or disorder is obesity, a metabolic disorder, a cardiovascular disorder, diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), or a combination of any of the foregoing. For example, in several embodiments, the FADS1-mediated disease or disorder is obesity. In several embodiments, the FADS1-mediated disease or disorder is a metabolic disorder. In several embodiments, the FADS1-mediated disease or disorder is a cardiovascular disorder. In several embodiments, the FADS1-mediated disease or disorder is diabetes. In several embodiments, the FADS1-mediated disease or disorder is dyslipidemia. In several embodiments, the FADS1-mediated disease or disorder is non-alcoholic steatohepatitis (NASH).

[0158] Embodiment 52 provided herein is the method or compound of any one of embodiments 1 to 51, wherein the reference level of the biological marker is the average amount of the biological marker in a population of healthy subjects.

[0159] Embodiment 53 provided herein is the method or compound of any one of embodiments 1 to 52, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects not suffering from a FADS1 -mediated disease.

[0160] Embodiment 54 provided herein is the method or compound of any one of embodiments 1 to 53, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects having a body mass index (BMI) greater than or equal to 25.0.

[0161] Embodiment 55 provided herein is the method or compound of any one of embodiments 1 to 53, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects having a body mass index (BMI) greater than or equal to 30.0.

[0162] Embodiment 56 provided herein is the method or compound of any one of embodiments 1 to 55, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects with a body fat percentage equal to or less than 19%.

[0163] Embodiment 57 provided herein is the method or compound of embodiment 56, wherein the reference level of the biological marker is from a male population and wherein the subject is male.

[0164] Embodiment 58 provided herein is the method or compound of any one of embodiments 1 to 55, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects having a body fat percentage equal to or less than 32%.

[0165] Embodiment 59 provided herein is the method or compound of embodiment 58, wherein the reference level of the biological marker is from a female population and wherein the subject is female.

[0166] Embodiment 60 provided herein is a method or compound of any one of embodiments 1 to 59, wherein the level of a biological marker for a FADS1-mediated disease from the subject is quantified using a sample collected from the subject; wherein the sample is blood, plasma, or a tissue biopsy. For example, in several embodiments, the level of a biological marker for a FADS1-mediated disease from the subject is determined using a blood sample from the subject. In several embodiments, the level of a biological marker for a FADS1-mediated disease from the subject is determined using a plasma sample from the subject. In several embodiments, the level of a biological marker for a FADS1-mediated disease from the subject is determined using a tissue sample from the subject. In several embodiments, the tissue sample is collected as a biopsy. In several embodiments, the blood or plasma sample is collected using a needle.

[0167] Embodiment 61 provided herein is the method or compound of embodiment 60, wherein the tissue is adipose tissue or organ tissue.For example, in several embodiments, an adipose tissue sample is obtained from the subject.

[0168] As disclosed elsewhere herein, several embodiments relate to methods for determining and / or predicting whether a subject is a candidate for (e.g., will respond to) treatment with a FADS1 inhibitor compound. In several embodiments, whether a subject is a candidate for treatment is related to the activity of the FADS1 enzyme in the subject. As disclosed elsewhere herein, when FADS1 activity in a subject is increased relative to healthy subjects and / or relative to a subject lacking a FADS1-mediated condition (e.g., a disease or disorder), treatment with a FADS1 inhibitor may be beneficial. In several embodiments, methods for identifying a subject with FADS1 activity are provided. In several embodiments, methods for measuring or approximating the level of FADS1 activity in a subject are provided. In several embodiments, methods for determining whether a subject is in need of treatment with a FADS1 inhibitor are provided, as disclosed elsewhere herein. In several embodiments, identifying a subject with a FADS1-mediated condition, identifying a subject with increased FADS1 activity (relative to healthy subjects), and / or identifying a subject in need of treatment with a FADS1 inhibitor compound can be performed by analyzing the relative abundance and / or ratio of one or more biological indicators (e.g., biomarkers) of a FADS1-mediated disease or disorder in the subject. Thus, provided herein are biological markers (eg, biomarkers) to be measured and / or compared. Several embodiments disclosed herein relate to methods of selecting a subject for treatment with a FADS1 inhibitor compound based on the presence or relative abundance of one or more biological markers.

[0169] Advantageously, based on individual patient information and characteristics, a treatment regimen can be customized for that individual. For example, the administration of a FADS1 inhibitor compound may or may not be initiated based on the levels of biological indicators in the patient. Because healthy subjects have one or more biological indicators that differ in relative abundance (e.g., decrease or increase) when compared to subjects who need to be treated with a FADS1 inhibitor compound (e.g., treatment candidates), a patient selection step can be performed by measuring the biological indicators in the treatment candidates. During the selection step, the levels of biological indicators in the treatment candidates can be compared with the levels of biological indicators from healthy subjects or with the levels of biological indicators from other successfully treated subjects (e.g., before their treatment). When the relative abundance of these biological indicators indicates that the subject may or will benefit from treatment (e.g., with a FADS1 inhibitor compound) or when a FADS1 disease or disorder exists or is likely to occur, the subject can be selected for treatment.

[0170] Once a treatment is selected, a FADS1 inhibitor compound can be administered to the patient as disclosed elsewhere herein. If the levels of these biological indicators indicate that the subject will not benefit from treatment or that treatment success is unlikely, the subject can be excluded from treatment.

[0171] As disclosed elsewhere herein, in several embodiments, biological indicators can be extracted from samples collected from subjects before, during, and / or after treatment or measured therein. In several embodiments, the sample is a tissue sample (e.g., from the liver, an organ, or blood). In several embodiments, the sample is collected by biopsy or using a syringe (e.g., to collect blood or plasma samples). In several embodiments, the sample is a body fluid (e.g., urine, saliva, plasma, etc.).

[0172] In several embodiments, the relative abundance of one, two, three, four, five, six, seven, eight or more biological indicators can be used to assess whether a subject has increased FADS1 activity and / or is a candidate for treatment with a FADS1 inhibitor compound. Any combination of the different biological indicators disclosed herein can be used (e.g., in a patient selection step).

[0173] In some embodiments, the biological marker may include one or more SNPs in the FADS1 gene. In some embodiments, the subject in need of treatment is a subject lacking the rs174556 and / or rs7115739 SNPs of FADS1.

[0174] As disclosed elsewhere herein, in several embodiments, the biological indicator, or at least one biological indicator, is a ratio of PUFAs in a subject. In several embodiments, the ratio of PUFAs comprises or consists of an AA to DGLA ratio. In several embodiments, for a subject in need of treatment, the AA to DGLA ratio is equal to or at least about: 4:1, 9:2, 5:1, 11:2, 6:1, 7:1, 8:1, 10:1, or a range including and / or spanning the aforementioned values. In several embodiments, for a subject in need of treatment, the AA to DGLA ratio is about or at least about: 10%, 20%, 30%, 40%, 50% greater than the ratio in a healthy individual, or a range including and / or spanning the aforementioned values. For example, if a healthy individual has an AA to DGLA ratio of 4, a value 10% greater is 4.4.

[0175] As disclosed elsewhere herein, in several embodiments, biological indicators or at least one biological indicator are one or more cell types. In several embodiments, cell types include adipocytes (Adipo), B cells (Bcell), endothelial cells (Endo), hepatocytes (Hep), Kupffer cells (Kupff), bone marrow cells (Myel), natural killer cells (NK), T cells (Tcell) or any of the foregoing combinations. In several embodiments, the relative abundance of at least one cell type of the subject requiring treatment increases (for example, relative to healthy subjects and / or subjects not requiring treatment). In several embodiments, the relative abundance of cell types (for example, from samples) increases by equal to or at least about: 10%, 20%, 30%, 50%, 75%, 100%, 150%, 200%, 250%, 300%, 350%, 400%, 500% or include and / or span the scope of the above values. To illustrate this point, when relative abundance increases by 200%, i.e., by 2 times, relative abundance is twice as high. In some embodiments, the relative abundance of a cell type (e.g., from a sample) increases by or is equal to or at least about: 1.1-fold, 1.2-fold, 1.3-fold, 1.5-fold, 1.75-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold, 40-fold, or a range including and / or spanning the above values. In some embodiments, the relative abundance of at least one cell type in a subject decreases. In some embodiments, the relative abundance of a cell type (e.g., from a sample) decreases by or is equal to or at least about: 2.5%, 5%, 10%, 20%, 30%, 50%, 75%, 90%, 92.5%, 95%, 97.5%, 99%, 99.5%, or a range including and / or spanning the above values. In several embodiments, the relative abundance of a cell type (e.g., from a sample) is reduced by equal to or at least about: 1.5-fold, 2-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold, 40-fold, or a range including and / or spanning the above values. To illustrate, when the relative abundance is reduced by 90%, i.e., by 10-fold, then the relative abundance is ten times less (because the relative abundance is 10% of the reference abundance).

[0176] As disclosed elsewhere herein, in several embodiments, the biological marker or at least one biological marker is one or more metabolites. In several embodiments, these metabolites include one or more of the following: plasma cholesterol, free cholesterol, total cholesterol, cholesterol esters, malic acid, α-ketoglutaric acid, mannose, glucose, erythro-dihydrosphingosine, 5-O-methylsphingosine, threo-sphingosine, 1-hydroxy-2-amino-(cis, trans)-3,5-octadecadiene, 4-hydroxydihydrosphingosine, thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxy heptadecatrienoic acid, 14,15-dihydroxy In some embodiments, the relative abundance of at least one metabolite in a subject is increased (e.g., relative to healthy subjects and / or subjects not requiring treatment). In some embodiments, the relative abundance of a metabolite (e.g., from a sample) increases by or is equal to or at least about: 10%, 20%, 30%, 50%, 75%, 100%, 200%, 300%, 500%, 700%, 1000%, or a range including and / or spanning the above values. In some embodiments, the relative abundance of a metabolite (e.g., from a sample) increases by or is equal to or at least about: 1.1-fold, 1.2-fold, 1.3-fold, 1.5-fold, 1.75-fold, 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 7-fold, 10-fold, 20-fold, or a range including and / or spanning the above values. In some embodiments, the relative abundance of at least one metabolite in a subject decreases. In some embodiments, the relative abundance of a metabolite (e.g., from a sample) decreases by or is equal to or at least about: 10%, 20%, 30%, 50%, 75%, 80%, 90%, 95%, or a range including and / or spanning the above values. In several embodiments, the relative abundance of a metabolite (e.g., from a sample) is reduced by equal to or at least about: 1.5-fold, 2-fold, 4-fold, 5-fold, 8-fold, 10-fold, 20-fold, or a range including and / or spanning the above values.

[0177] As disclosed elsewhere herein, in several embodiments, the biological indicator or at least one biological indicator is one or more DEGs. In several embodiments, the DEGs include one or more of the following: Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Hsd11b1, Akr1d1, Aldh1a1, Mmp19, Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel113, Agap2, 4833411C07Rik, Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Ta gln2, Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, Cyp2b9, Fads1, Hao2, Cyp2b13, Cyp2a22 , Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, Fitm1, Tmem86a, Agap2, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1 , A730063M14Rik, Ly6f, Foxi1, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, Ppp2r5b, Slc6a7 , Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Mfap2, Gm18537, Aacs, Pclo, Adrb3, Gm38394, AC154232.2. Cadps, Adgrb2, Gm45470, Sdr9c7, Dsg1c, Slc17a1, Acnat2, Ces1c, Gss, Hsd11b1, 1810008I18Rik, Tlcd1, Snrk, Akr1c20 , Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acss2, Acacb, Arhgap27, Adam7, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl 19. Tcf7, ,1600015I10Rik,Sez6I2,Prnd,Gm16702,S100a8,Pcdh12,Malat1,Kcnq1ot1,ArI4c,Gm38394,Gm42549,AC154232.2,Gm37 310, Gm37776, Atp2a1, Ckm, Tnnt3, Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Hsd11b1, Akr1d1, Aldh1a1, Mmp19, A dipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9, Cebpa, Dgat1, Dgat2, Elovl3, Fads1, Fads2, Fas, Scd1, Srebf1, Adrb3, Hilpda, Li pe, Mgll, Plin1, Plin4, Pnpla2, Pnpla3, Ldah, Cs, Gckr, Mel, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, Tm6sf2, Tnf, or any combination of the foregoing. In several embodiments, the DEGs include one or more of those shown in any one or more of Figures 4a, 5a, 10a, 13a, 13g, 15e, and 15f, or a combination of the DEGs disclosed above. In several embodiments, the DEGs are measured using RNA sequence analysis as disclosed elsewhere herein.

[0178] In several embodiments, the relative abundance of at least one DEG of a subject increases (e.g., relative to healthy subjects and / or subjects not requiring treatment). In several embodiments, the relative abundance of a DEG (e.g., from a sample) increases by an amount equal to or at least about: 2 times, 3 times, 4 times, 5 times, 10 times, 20 times, 30 times, 40 times, 50 times, 75 times, 100 times, 150 times, 200 times, 1000 times, 100,000 times, 200,000 times or including and / or spanning the scope of the above values. In several embodiments, the relative abundance of at least one DEG of a subject decreases. In several embodiments, the relative abundance of a DEG (e.g., from a sample) is reduced by equal to or at least about: 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 75-fold, 100-fold, 150-fold, 200-fold, 1000-fold, 100,000-fold, 200,000-fold, or a range including and / or spanning the above values.

[0179] As disclosed elsewhere herein, several embodiments provide methods of treatment. In several embodiments, the subject to be treated suffers from a FADS1-mediated disease or disorder (e.g., a disease or disorder that can be treated by regulating the FADS1 enzyme). In several embodiments, a biological indicator of FADS1 activity is measured in the subject before, simultaneously with, or after administration of a FADS1 inhibitor compound. In several embodiments, the FADS1-mediated disease or disorder is one or more of increased body mass index, obesity, metabolic disorders, cardiovascular disorders, diabetes, dyslipidemia, and / or non-alcoholic steatohepatitis (NASH). The scope of the methods and uses provided in this disclosure should be understood to encompass methods and uses employing all compounds disclosed herein. In addition to being useful for human treatment, the compounds provided herein can also be used in veterinary treatment of companion animals, exotic animals, and farm animals, including mammals, rodents, and the like. For example, animals including horses, dogs, and cats can be treated with the compounds provided herein.

[0180] In several embodiments, a compound disclosed herein or a pharmaceutical composition comprising the compound is provided for reducing the weight of a subject. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for reducing the body mass index of a subject. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating metabolic disorders. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating cardiovascular disorders. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating diabetes. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating obesity. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating dyslipidemia. In several embodiments, a compound or a pharmaceutical composition comprising the compound is provided for treating non-alcoholic steatohepatitis (NASH).

[0181] In some embodiments, the compound or a pharmaceutical composition comprising the compound is used to prepare a medicament for reducing the weight or body mass index of a subject. In some embodiments, the compound or a pharmaceutical composition comprising the compound is used to prepare a medicament for treating a metabolic or cardiovascular disorder. In some embodiments, the compound or a pharmaceutical composition comprising the compound is used to prepare a medicament for treating diabetes, obesity, dyslipidemia, or non-alcoholic steatohepatitis (NASH).

[0182] In several embodiments, the compound or a pharmaceutical composition comprising the compound is used in a method of reducing the weight and / or body mass index of a subject in need thereof. In several embodiments, the compound or a pharmaceutical composition comprising the compound is used in a method of treating a metabolic and / or cardiovascular disorder in a subject in need thereof. In several embodiments, the compound or a pharmaceutical composition comprising the compound is used in a method of treating diabetes, obesity, dyslipidemia, and / or non-alcoholic steatohepatitis (NASH) in a subject in need thereof. In several embodiments, the compound or a pharmaceutical composition comprising the compound is used in a method of reducing waist-to-hip ratio (WHR) in a subject in need thereof. In several embodiments, the method comprises administering to the subject a therapeutically effective amount of the compound or composition.

[0183] Another embodiment provided herein is a method for lowering blood sugar in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising such a compound. In several embodiments, the method lowers blood sugar by 10% or more. In several embodiments, the method lowers blood sugar by 15% or more. In several embodiments, the method lowers blood sugar by 20% or more. In several embodiments, the method lowers blood sugar by 25% or more. In several embodiments, the method lowers blood sugar by 30% or more. In several embodiments, the method lowers blood sugar by 35% or more. In several embodiments, the method lowers blood sugar by 40% or more. In several embodiments, the method lowers blood sugar by 50% or more. In several embodiments, the method lowers blood sugar with minimal effect on food intake / appetite. In several embodiments, the method lowers blood sugar with no effect on food intake / appetite.

[0184] Another embodiment provided herein is a method of reducing insulin in a subject in need thereof, the method comprising administering to the subject a compound or a pharmaceutical composition comprising such a compound. In several embodiments, the method reduces insulin by 50% or more. In several embodiments, the method reduces insulin by 60% or more. In several embodiments, the method reduces insulin by 70% or more. In several embodiments, the method reduces insulin by 80% or more. In several embodiments, the method reduces blood insulin by 85% or more. In several embodiments, the method reduces insulin by 86% or more. In several embodiments, the method reduces insulin by 87% or more. In several embodiments, the method reduces insulin by 88% or more. In several embodiments, the method reduces insulin by 89% or more. In several embodiments, the method reduces insulin by 90% or more. In several embodiments, the method reduces insulin by 91% or more. In several embodiments, the method reduces insulin with minimal effect on food intake / appetite. In several embodiments, the method reduces insulin with no effect on food intake / appetite.

[0185] Another embodiment provided herein is a method for lowering cholesterol in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising such a compound. In several embodiments, the method reduces cholesterol by 10% or more. In several embodiments, the method reduces cholesterol by 15% or more. In several embodiments, the method reduces cholesterol by 20% or more. In several embodiments, the method reduces cholesterol by 30% or more. In several embodiments, the method reduces cholesterol by 31% or more. In several embodiments, the method reduces cholesterol by 32% or more. In several embodiments, the method reduces cholesterol by 33% or more. In several embodiments, the method reduces cholesterol by 34% or more. In several embodiments, the method reduces cholesterol by 35% or more. In several embodiments, the method reduces blood cholesterol by 36% or more. In several embodiments, the method reduces cholesterol by 37% or more. In several embodiments, the method reduces cholesterol by 38% or more. In several embodiments, the method reduces cholesterol by 39% or more. In several embodiments, the method reduces cholesterol with minimal effect on food intake / appetite. In several embodiments, the method reduces cholesterol with no effect on food intake / appetite.

[0186] Another embodiment provided herein is a method for lowering LDL in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising the compound. In several embodiments, the method reduces low-density lipoprotein (LDL) by 10% or more. In several embodiments, the method reduces LDL by 20% or more. In several embodiments, the method reduces LDL by 21% or more. In several embodiments, the method reduces LDL by 22% or more. In several embodiments, the method reduces LDL by 23% or more. In several embodiments, the method reduces LDL by 24% or more. In several embodiments, the method reduces LDL by 25% or more. In several embodiments, the method reduces LDL by 26% or more. In several embodiments, the method reduces blood LDL by 27% or more. In several embodiments, the method reduces LDL while having minimal effect on food intake / appetite. In several embodiments, the method reduces LDL while having no effect on food intake / appetite.

[0187] Another embodiment provided herein is a method of lowering triglycerides in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising such a compound. In several embodiments, the method reduces triglycerides by 30% or more. In several embodiments, the method reduces triglycerides by 40% or more. In several embodiments, the method reduces triglycerides by 50% or more. In several embodiments, the method reduces triglycerides by 51% or more. In several embodiments, the method reduces triglycerides by 52% or more. In several embodiments, the method reduces triglycerides by 53% or more. In several embodiments, the method reduces triglycerides by 54% or more. In several embodiments, the method reduces triglycerides by 55% or more. In several embodiments, the method reduces blood triglycerides by 56% or more. In several embodiments, the method reduces triglycerides by 57% or more. In several embodiments, the method reduces triglycerides while having minimal effect on food intake / appetite. In several embodiments, the method reduces triglycerides while having no effect on food intake / appetite

[0188] Another embodiment provided herein is a method for reducing fat mass in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising the compound. In several embodiments, the method reduces fat mass in the subject by 30% or more. In several embodiments, the method reduces fat mass in the subject by 40% or more. In several embodiments, the method reduces fat mass in the subject by 45% or more. In several embodiments, the method reduces fat mass in the subject by 50% or more. In several embodiments, the method reduces fat mass in the subject by 55% or more. In several embodiments, the method reduces blood fat levels in the subject by 60% or more. In several embodiments, the method reduces fat mass in the subject by 65% or more. In several embodiments, the method reduces fat mass in the subject by 70% or more. In several embodiments, the method reduces fat mass in the subject by 75% or more. In several embodiments, the method reduces fat mass in the subject while having minimal effect on food intake / appetite. In several embodiments, the method reduces fat mass in the subject while having no effect on food intake / appetite.

[0189] Another embodiment provided herein is a method of increasing adiponectin in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising the compound.

[0190] Another embodiment provided herein is a method of lowering leptin in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising the compound.

[0191] Another embodiment provided herein is a method of reducing resistance in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutical composition comprising the compound.

[0192] The various method steps described above may be performed in an alternate order or sequence to give the desired results.

[0193] In addition, the treatment results of the methods disclosed herein provide further opportunities to customize dosing regimens for patients. For example, if the treatment results (e.g., weight loss, reduction in dyslipidemia, etc.) after a period of treatment (e.g., equal to or greater than 2 months, 6 months, etc.) are lower than expected, the dosage of the FADS1 inhibitor may be increased. When the downstream treatment results (e.g., weight loss, reduction in dyslipidemia, etc.) after a period of treatment (e.g., equal to or greater than 2 months, 6 months, etc.) are higher than expected or desired, the dosage of the FADS1 inhibitor may be maintained or reduced.

[0194] FADS1 inhibitor compounds

[0195] In several embodiments, methods of treating a FADS1-mediated disease or disorder are performed by administering to a subject in need of treatment a compound disclosed herein (e.g., a FADS1 inhibitory compound), a tautomer thereof, or a pharmaceutically acceptable salt of the compound or tautomer.

[0196] Embodiment 62 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0197] 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0198] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0199] 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione;

[0200] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0201] 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0202] 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-3,7-bis(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0203] 2-Fluoro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0204] 7-(Trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0205] 2-chloro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0206] 2-(methoxymethyl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0207] 2-cyclopropyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0208] 2-cyclopropyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0209] 2,3-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one;

[0210] 2,3-Dimethyl-5-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one;

[0211] 7-ethyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0212] 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one;

[0213] 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one;

[0214] 1,3-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one;

[0215] 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,8H-pyrrolo[1,2-a]pyrimidin-4-one;

[0216] 2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0217] 2-(Trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H,6H,7H,8H-pyrrolo[1,2-a]pyrimidin-4-one;

[0218] 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0219] 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0220] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0221] 8-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-9-(trifluoromethyl)-6,10-diazatricyclo[4.4.0.0 2,4 ]Deca-1(10),8-dien-7-one;

[0222] 6-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0223] 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-[1,2,4]triazolo[4,3-a]pyrimidin-5-one;

[0224] 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,3,4]tetrazo[1,5-a]pyrimidin-7-one;

[0225] 2-methyl-6-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0226] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0227] 6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0228] 2-(Hydroxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0229] 2-(Hydroxymethyl)-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0230] 2-chloro-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0231] 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(propan-2-yl)-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione;

[0232] 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0233] 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one;

[0234] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]oxazolo[3,2-a]pyrimidin-5-one;

[0235] 2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0236] 7-Ethoxy-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0237] 2-(methoxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0238] 2-methoxy-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0239] 3-chloro-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0240] 2-(Hydroxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0241] 2-(Hydroxymethyl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0242] 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione;

[0243] 2-chloro-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0244] 2-chloro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0245] 2-cyclopropyl-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0246] 2-chloro-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0247] 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0248] 1,2-dimethyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0249] 1,2-Dimethyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0250] 2-(methoxymethyl)-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0251] 1-ethyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0252] 1-(2-methoxyethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0253] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(propan-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0254] 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0255] 6-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0256] 6-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0257] 1-(cyclopropylmethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0258] 2-(methoxymethyl)-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0259] 1-(2-Hydroxypropyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0260] 1,2-dimethyl-6-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0261] 1-(cyclopropylmethyl)-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0262] 1-[2-(dimethylamino)ethyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0263] 1-(cyclopropylmethyl)-2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0264] 1-[2-(dimethylamino)ethyl]-2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0265] 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0266] 2-methoxy-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0267] 2-methoxy-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0268] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0269] 2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0270] 2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0271] 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0272] 1-( 2 H3) methyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0273] 1-( 2 H3) methyl-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0274] 1-(2-hydroxyethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0275] 2-methyl-5-oxo-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidine-1-carboxylic acid methyl ester;

[0276] 1-[(2,2-difluorocyclopropyl)methyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0277] 1-[(3,3-difluorocyclobutyl)methyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0278] 1-(2-hydroxyethyl)-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0279] 1-[2-(dimethylamino)ethyl]-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0280] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(prop-2-yn-1-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0281] 2-{2-methyl-5-oxo-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-1-yl}acetonitrile;

[0282] 2-[2-methyl-5-oxo-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-1-yl]acetonitrile;

[0283] 1-(2-hydroxy-2-methylpropyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0284] 1-[2-(1-hydroxycyclopropyl)ethyl]-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0285] 2-methyl-1-[(oxetan-3-yl)methyl]-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0286] 2-methyl-1-(oxetan-3-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0287] 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidine-5-thione;

[0288] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0289] 2-methyl-1-(pyridin-2-yl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0290] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyrazin-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0291] 2-methyl-1-(6-methylpyridin-2-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0292] 2-methyl-1-(1-methyl-1H-pyrazol-4-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0293] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-3-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0294] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-phenyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0295] 1-(6-chloropyridin-2-yl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0296] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-4-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0297] 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(1H-pyrazol-4-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0298] 2-(Fluoromethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0299] 2-[(dimethylamino)methyl]-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0300] 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0301] 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0302] 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0303] 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0304] 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0305] 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one;

[0306] (2R)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0307] (2S)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H,6H,7H-[1,3]thiazolo[3,2-a]pyrimidin-5-one;

[0308] 1-{[(1R)-2,2-difluorocyclopropyl]methyl}-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0309] 1-{[(1S)-2,2-difluorocyclopropyl]methyl}-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one;

[0310] (2R)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0311] (2R)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0312] (2S)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0313] (2S)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0314] (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)-1H-pyrazol-1-yl)acetonitrile;

[0315] (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)acetonitrile;

[0316] (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenyl)acetonitrile;

[0317] (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)acetonitrile;

[0318] (4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetonitrile;

[0319] 1-(Chloromethyl)-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-pyrimido[1,2-a][1,3]diazine-2,6-dione;

[0320] 1-(Fluoromethyl)-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione;

[0321] 1-(methyl-d3)-7-(4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione;

[0322] 1-Methyl-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione;

[0323] 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)-2-methylpropionitrile;

[0324] 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0325] 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile;

[0326] 2-(difluoromethyl)-3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-4H-pyrido[1,2-a]pyrimidin-4-one;

[0327] 2-(Difluoromethyl)-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile;

[0328] 2-(difluoromethyl)-8-methoxy-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0329] 2-(difluoromethyl)-8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0330] 2-(Difluoromethyl)-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0331] 2-(difluoromethyl)-8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0332] 2-(Fluoromethyl)-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile;

[0333] 2-(Fluoromethyl)-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0334] 2-(Trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H,6H,7H,9H-pyrimido[2,1-c][1,4]oxazin-4-one;

[0335] 2,8-dimethoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0336] 2-cyclopropyl-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0337] 2-Ethoxy-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0338] 2-Ethoxy-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0339] 2-ethyl-8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0340] 2-ethyl-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0341] 2-ethyl-8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0342] 3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-4,8(1H)-dione;

[0343] 3-(1-(2,2-difluoropropyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0344] 3-(1-(3-fluorophenyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0345] 3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0346] 3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0347] 3-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0348] 3-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0349] 3-(1-Benzofuran-2-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0350] 3-(1-cyclopropyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0351] 3-(1-cyclopropyl-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0352] 3-(1-phenyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0353] 3-(1-propyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0354] 3-(2-chloro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0355] 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0356] 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0357] 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0358] 3-(2-fluoro-4-(trifluoromethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0359] 3-(2-fluoro-6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0360] 3-(3-chloro-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0361] 3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0362] 3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0363] 3-(4-(((1R)-2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0364] 3-(4-(((1S)-2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0365] 3-(4-((2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0366] 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0367] 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-4,8(1H)-dione;

[0368] 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2,8-bis(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0369] 3-(4-(2,2-difluoroethoxy)-2-fluorophenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0370] 3-(4-(2,2-difluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0371] 3-(4-(2,2-difluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0372] 3-(4-(2,2-difluoropropoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0373] 3-(4-(2-fluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0374] 3-(4-(2-fluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0375] 3-(4-(2-fluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0376] 3-(4-(2-fluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0377] 3-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)-1H-pyrazol-1-yl)propionitrile;

[0378] 3-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenyl)propionitrile;

[0379] 3-(4-(cyclopropylmethoxy)-2-fluorophenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0380] 3-(4-(cyclopropylmethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0381] 3-(4-(cyclopropylmethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0382] 3-(4-(cyclopropylmethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0383] 3-(4-(difluoromethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0384] 3-(5-(2,2,2-trifluoroethoxy)-2-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0385] 3-(5-fluoro-6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0386] 3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0387] 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,9H-pyrimido[2,1-c][1,4]oxazin-4-one;

[0388] 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0389] 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0390] 3-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0391] 3-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0392] 3-[5-iodo-1-(2,2,3,3,3-pentafluoropropyl)-1H-1,2,3-triazol-4-yl]-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0393] 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-7-fluoro-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0394] 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0395] 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0396] 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0397] 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0398] 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0399] 3-Fluoro-1-methyl-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione;

[0400] 4-Oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile;

[0401] 4-Oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylic acid;

[0402] 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile;

[0403] 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide;

[0404] 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylic acid;

[0405] 4-Oxo-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-7-carbonitrile;

[0406] 7-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione;

[0407] 7-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione;

[0408] 7-(4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione;

[0409] 7-(4-(2-fluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione;

[0410] 7-(methoxymethyl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0411] 7,8-dimethyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one;

[0412] 7,8-dimethyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0413] 7,9-dimethyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0414] 7-chloro-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0415] 7-chloro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0416] 7-chloro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0417] 7-chloro-8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0418] 7-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one;

[0419] 7-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0420] 7-chloro-8-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0421] 7-chloro-8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0422] 7-chloro-8-methyl-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0423] 7-cyclopropyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0424] 7-cyclopropyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0425] 7-Fluoro-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0426] 7-Fluoro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0427] 7-Fluoro-8-hydroxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0428] 7-Fluoro-8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0429] 7-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one;

[0430] 7-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0431] 7-Fluoro-8-methoxy-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0432] 7-Fluoro-8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0433] 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one;

[0434] 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0435] 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0436] 7-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one;

[0437] 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one;

[0438] 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0439] 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0440] 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0441] 8-((1R)-1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0442] 8-((1R)-1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0443] 8-((1S)-1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0444] 8-((1S)-1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0445] 8-((dimethylamino)methyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0446] 8-(methyloxy-d3)-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0447] 8-((methylsulfanyl)methoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0448] 8-((R)-ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0449] 8-((R)-methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0450] 8-((R)-methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0451] 8-((S)-ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0452] 8-((S)-methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0453] 8-((S)-methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0454] 8-(1,3-oxazol-2-yl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0455] 8-(1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0456] 8-(1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0457] 8-(2-hydroxypropan-2-yl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0458] 8-(2-methyl-2-oxetanyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0459] 8-(2-propyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0460] 8-(3-azetidinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0461] 8-(aminomethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0462] 8-(1-azetidinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0463] 8-(Chloromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0464] 8-(Chloromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0465] 8-(difluoromethoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0466] 8-(difluoromethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0467] 8-(dimethylamino)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0468] 8-(dimethylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0469] 8-(ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0470] 8-(Fluoromethoxy)-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0471] 8-(Fluoromethoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0472] 8-(Fluoromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0473] 8-(Fluoromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0474] 8-(Fluoromethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0475] 8-(Fluoromethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0476] 8-(Hydroxymethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0477] 8-(Hydroxymethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0478] 8-(methoxymethyl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0479] 8-(methylamino)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0480] 8-(methylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0481] 8-(methylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0482] 8-(methyl-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0483] 8-(methyloxy-d3)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0484] 8-(methyloxy-d3)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0485] 8-(methyloxy-d3)-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0486] 8-(methyloxy-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0487] 8-(methyloxy-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0488] 8-(methyloxy-d3)-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0489] 8-(Methylsulfanyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0490] 8-(Methylsulfanyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0491] 8-(methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0492] 8-(methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0493] 8-(methylsulfonyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0494] 8-acetyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0495] 8-amino-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0496] 8-amino-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0497] 8-amino-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0498] 8-chloro-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0499] 8-chloro-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0500] 8-cyclopropyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0501] 8-cyclopropyl-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0502] 8-vinyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0503] 8-Ethoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0504] 8-ethyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0505] 8-fluoro-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0506] 8-Hydroxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0507] 8-methoxy-2-(trifluoromethyl)-3-(1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0508] 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0509] 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0510] 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0511] 8-methoxy-2-(trifluoromethyl)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0512] 8-methoxy-2-(trifluoromethyl)-3-(4-(3,3,3-trifluoropropyl)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0513] 8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one;

[0514] 8-methoxy-2-(trifluoromethyl)-3-[3-(3,3,3-trifluoropropyl)-1,2-oxazol-5-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0515] 8-methoxy-2-(trifluoromethyl)-3-[4-(3,3,3-trifluoropropyl)-1H-imidazol-1-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0516] 8-methoxy-2-(trifluoromethyl)-3-[5-(3,3,3-trifluoropropyl)-1,3-thiazol-2-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0517] 8-methoxy-2-methyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0518] 8-methoxy-3-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0519] 8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0520] 8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0521] 8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0522] 8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0523] 8-methoxy-3-(1-phenyl-1H-pyrazol-3-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0524] 8-methoxy-3-(1-phenyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0525] 8-methoxy-3-(1-propyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0526] 8-methoxy-3-(2-methyl-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0527] 8-methoxy-3-(2-phenyl-1,3-oxazol-5-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0528] 8-methoxy-3-(3-phenyl-1,2-oxazol-5-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0529] 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)-2-(trifluoromethyl)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0530] 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0531] 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0532] 8-methoxy-3-(4-(2,2,3,3,3-pentafluoropropoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0533] 8-methoxy-3-(4-(2,2,3,3-tetrafluoropropoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0534] 8-methoxy-3-(4-(trifluoromethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0535] 8-methoxy-3-(4-(trifluoromethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0536] 8-methoxy-3-(4-propylphenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0537] 8-methoxy-3-(5-propyl-1,2-oxazol-3-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0538] 8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0539] 8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one;

[0540] 8-methoxy-3-(6-propyl-3-pyridyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0541] 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-1,2,3-triazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0542] 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0543] 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,6-a]pyrimidin-4-one;

[0544] 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0545] 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)-1,3-thiazol-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0546] 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one;

[0547] 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0548] 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0549] 8-methoxy-3-[2-(2,2,3,3,3-pentafluoropropoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0550] 8-methoxy-3-[3-(2,2,3,3,3-pentafluoropropyl)-1,2-oxazol-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0551] 8-methoxy-3-[4-(2,2,2-trifluoroethoxy)-1,3-thiazol-2-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0552] 8-methoxy-3-[5-(2,2,2-trifluoroethoxy)-1,3-thiazol-2-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0553] 8-methoxy-3-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0554] 8-methoxy-6-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0555] 8-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one;

[0556] 8-methyl-2-(trifluoromethyl)-3-[5-(3,3,3-trifluoropropyl)-1,2,4-oxadiazol-3-yl]-4H-pyrido[1,2-a]pyrimidin-4-one;

[0557] 8-methyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0558] 8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,8H,9H-pyrimido[1,2-a]pyrazin-4-one;

[0559] 8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0560] 8-methyl-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0561] 8-methyl-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one;

[0562] 9-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0563] 9-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one;

[0564] 9-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one;

[0565] Methyl 4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylate;

[0566] Methyl(4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)carbamoyl fluoride;

[0567] N-(4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetamide;

[0568] N-(4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetamide;

[0569] N,N-dimethyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide;

[0570] N-ethyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide;

[0571] N-methyl-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; or

[0572] N-methyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; or

[0573] A pharmaceutically acceptable salt of any of the foregoing.

[0574] Embodiment 63 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0575]

[0576] or a pharmaceutically acceptable salt thereof.

[0577] Embodiment 64 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0578]

[0579] or a pharmaceutically acceptable salt thereof.

[0580] Embodiment 65 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0581]

[0582] or a pharmaceutically acceptable salt thereof.

[0583] Embodiment 66 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0584]

[0585] or a pharmaceutically acceptable salt thereof.

[0586] Embodiment 67 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0587]

[0588] or a pharmaceutically acceptable salt thereof.

[0589] Embodiment 68 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0590]

[0591] or a pharmaceutically acceptable salt thereof.

[0592] Embodiment 69 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0593]

[0594] or a pharmaceutically acceptable salt thereof.

[0595] Embodiment 70 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0596]

[0597] or a pharmaceutically acceptable salt thereof.

[0598] Embodiment 71 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0599]

[0600] or a pharmaceutically acceptable salt thereof.

[0601] Embodiment 72 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0602]

[0603] or a pharmaceutically acceptable salt thereof.

[0604] Embodiment 73 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0605]

[0606] or a pharmaceutically acceptable salt thereof.

[0607] Embodiment 74 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0608]

[0609] or a pharmaceutically acceptable salt thereof.

[0610] Embodiment 75 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0611]

[0612] or a pharmaceutically acceptable salt thereof.

[0613] Embodiment 76 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0614]

[0615] or a pharmaceutically acceptable salt thereof.

[0616] Embodiment 77 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0617]

[0618] or a pharmaceutically acceptable salt thereof.

[0619] Embodiment 78 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0620]

[0621] or a pharmaceutically acceptable salt thereof.

[0622] Embodiment 79 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0623]

[0624] or a pharmaceutically acceptable salt thereof.

[0625] Embodiment 80 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0626]

[0627] or a pharmaceutically acceptable salt thereof.

[0628] Embodiment 81 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0629]

[0630] or a pharmaceutically acceptable salt thereof.

[0631] Embodiment 82 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0632]

[0633] or a pharmaceutically acceptable salt thereof.

[0634] Embodiment 83 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0635]

[0636] or a pharmaceutically acceptable salt thereof.

[0637] Embodiment 84 provided herein is the method or compound of any one of embodiments 1 to 61, wherein the FADS1 inhibitor compound is:

[0638]

[0639] or a pharmaceutically acceptable salt thereof.

[0640] Embodiment 85 provided herein is the method or compound of any one of claims 1 to 84, wherein the FADS1 inhibitor compound is the free base.

[0641] Patient Response and Labeled DGLA

[0642] Also disclosed herein are methods for evaluating, analyzing, and / or adjusting the dosing regimen of a patient who has received a FADS1 inhibitor compound treatment or is in an ongoing treatment regimen. Using one or more of the methods disclosed herein, the level of inhibition that occurs with a dose of a FADS1 inhibitor compound can be tested relative to the maximum inhibition level (e.g., IC level). When the inhibitory concentration level is below the target and / or desired level, the dose can be adjusted (e.g., upward). When the inhibitory concentration level is equal to or above the target level, the dose can be adjusted downward or maintained. For illustration, when the target therapeutic dose is an inhibitory concentration of 90% (IC90) of the maximum inhibitory concentration or above IC90 and the actual dose provides 50% inhibition (e.g., IC50), the dose of the FADS1 inhibitor can be increased (a larger dose or increased dose frequency). In some embodiments, the target therapeutic inhibitory concentration is equal to or at least about: 70% (e.g., IC70), 80% (e.g., IC80), 90% (e.g., IC90), 95% (e.g., IC95), 97.5% (e.g., IC97.5), 99% (e.g., IC99), 99.9% (e.g., IC99.9), 100% (IC100), or a range including and / or spanning the above values. In some embodiments, the measured and / or actual inhibitory concentration at the administered dose is equal to or less than about: 10% (e.g., IC10), 20% (e.g., IC20), 30% (e.g., IC30), 40% (e.g., IC40), 50% (e.g., IC50), 60% (e.g., IC60), 70% (e.g., IC70), 80% (e.g., IC80), 90% (e.g., IC90), or a range including and / or spanning the above values. In some embodiments, where the actual inhibitory concentration is equal to or less than about: 10% (e.g., IC10), 20% (e.g., IC20), 30% (e.g., IC30), 50% (e.g., IC50), 60% (e.g., IC60), 70% (e.g., IC70), 80% (e.g., IC80), 90% (e.g., IC90), 95% (e.g., IC95), or a range including and / or spanning the above values for the maximum inhibitory concentration, the dose can be increased. In some embodiments, when the inhibitory concentration is lower than desired, the dose is increased by about or at least about: 5%, 10%, 15%, 20%, 30%, 50%, 75%, 100%, 200%, or a range including and / or spanning the above values. In some embodiments, when the inhibitory concentration is lower than desired, the dosing frequency is doubled. In some embodiments, both the dose and the dose frequency can be increased.

[0643] In several embodiments, to measure the inhibitory concentration achieved with a specific FADS1 inhibitory compound, a labeled DGLA molecule can be administered to a subject (e.g., orally, intravascularly, intravenously, intraarterially, intraperitoneally, subcutaneously, etc.). After a period of time, a sample is collected from the subject, as disclosed elsewhere herein. Thereafter, the amount of labeled AA and the amount of labeled DGLA in the sample can be determined. The values for labeled AA and labeled DGLA can be used to calculate the inhibitory concentration (e.g., using the methods disclosed in the Examples section). In several embodiments, the sample is a tissue biopsy or a blood sample (e.g., a plasma sample). In several embodiments, the time period between administration of labeled DGLA and sample collection is equal to or at least about: 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 18 hours, 24 hours, or a range including and / or spanning the above values.

[0644] In some embodiments, a FADS1 inhibitor compound is administered to a subject prior to, concurrently with, or after administration of a labeled DGLA molecule (e.g., an isotopologue of DGLA). In some embodiments, the time period between administration of the FADS1 inhibitor and administration of labeled DGLA is equal to or at least about: 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 18 hours, 24 hours, or a range including and / or spanning the above values.

[0645] In several embodiments, the labeled DGLA comprises isotopically enriched at one or more positions 13 In some embodiments, the labeled DGLA comprises an isotopically enriched 13 In some embodiments, the labeled DGLA comprises DGLA having an isotopically enriched 13 In some embodiments, the labeled DGLA comprises DGLA having an isotopically enriched 13 In some embodiments, the labeled DGLA comprises DGLA having an isotopically enriched 13 In some embodiments, the labeled DGLA comprises isotopically enriched DGLA at the following positions: 13C atoms at positions equal to or at least about 1 position, 2 positions, 3 positions, 4 positions, 5 positions, 6 positions, 7 positions, 8 positions, 9 positions, 10 positions, 11 positions, 12 positions, 13 positions, 14 positions, 15 positions, 16 positions, 17 positions, 18 positions, 19 positions, 20 positions, or a range including and / or spanning the above values. In several embodiments, the labeled DGLA comprises a uniformly enriched isotope. 13 DGLA of C atoms.

[0646] In several embodiments, the labeled DGLA comprises the following structure (formula):

[0647]

[0648] Each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13 C. In several embodiments, the compound is referred to as 13 C w -DGLA, where "w" is the enriched isotope 13 The number of "*" positions of C. In several embodiments, "w" is equal to or at least 1, 2, 3, 4 or 5. For example, when w is 2, then the two "*" symbols represent the enriched isotope. 13 In some embodiments, equal to or at least two "*" symbols indicate enriched isotopes 13 In some embodiments, equal to or at least three "*" symbols indicate enriched isotopes 13 In some embodiments, equal to or at least four "*" symbols indicate enriched isotopes 13 In some embodiments, equal to or at least five "*" symbols indicate enriched isotopes 13 The position of C.

[0649] In several embodiments, the measurement of the biological indicators disclosed elsewhere herein can be incorporated into the methods of treatment and uses disclosed elsewhere herein. In several embodiments, the measurement of the biological indicators disclosed elsewhere herein and / or the patient selection steps are used to reduce weight, reduce body mass index, treat obesity, treat metabolic disorders, treat cardiovascular disorders, treat diabetes, treat obesity, treat dyslipidemia and / or treat non-alcoholic steatohepatitis ("NASH") methods. The methods and uses that can be combined with the measurement and selection steps described in this section are described elsewhere herein.

[0650] Example 86 provided herein is a method of measuring the ratio of arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA) in a subject;

[0651] wherein the AA is isotopically labeled and the DGLA is isotopically labeled;

[0652] Wherein the AA to DGLA ratio is measured by administering a dose of labeled DGLA to a subject and then measuring the ratio of labeled AA to labeled DGLA.

[0653] Example 87 provided herein is the method of Example 86, wherein the labeled DGLA comprises an isotopically enriched 13 DGLA of C.

[0654] Embodiment 88 provided herein is the method of embodiment 87, wherein the labeled DGLA comprises isotopically enriched 13 For example, in some embodiments, the labeled DGLA is isotopically enriched. 13 In some embodiments, the labeled DGLA is isotopically enriched. 13 DGLA at more than two carbon positions of C atoms.

[0655] Embodiment 89 provided herein is the method of embodiment 87, wherein the labeled DGLA comprises isotopically enriched 13 For example, in some embodiments, the labeled DGLA is isotopically enriched. 13 In some embodiments, the labeled DGLA is isotopically enriched. 13 DGLA with more than three carbon positions of C atoms.

[0656] Example 90 provided herein is the method of Example 87, wherein the labeled DGLA comprises isotopically enriched 13 For example, in some embodiments, the labeled DGLA is isotopically enriched. 13 In some embodiments, the labeled DGLA is isotopically enriched. 13 DGLA with more than four carbon positions of C atoms.

[0657] Embodiment 91 provided herein is the method of embodiment 87, wherein the labeled DGLA comprises a carbonyl group having equal to or at least five carbon position enriched isotopes. 13 For example, in some embodiments, the labeled DGLA is isotopically enriched. 13In some embodiments, the labeled DGLA is isotopically enriched. 13 DGLA with more than five carbon positions of C atoms.

[0658] Embodiment 92 provided herein is the method of any one of embodiments 86 to 91, wherein the labeled DGLA comprises a label at one or more or all of the carbons indicated below with "*":

[0659]

[0660] In some embodiments, equal to or at least two "*" symbols indicate isotope enrichment. 13 In some embodiments, equal to or at least three "*" symbols indicate enriched isotopes 13 In some embodiments, equal to or at least four "*" symbols indicate enriched isotopes 13 In several examples, each of the five "*" symbols represents an enriched isotope. 13 The position of C.

[0661] Embodiment 93 provided herein is the method of any one of embodiments 86 to 92, wherein a dose of a FADS1 inhibitor compound is administered to the subject prior to, simultaneously with, or after administration of the labeled dose of DGLA to the subject.

[0662] Example 94 provided herein is the method of Example 92, wherein the conversion of labeled DGLA to labeled AA is used to measure and / or calculate the level of inhibition of the FADS1 enzyme by the dose of the FADS1 inhibitor compound provided to the patient.

[0663] Example 95 provided herein is the method of Example 94, further comprising comparing the measured level of inhibition of the dose of the FADS1 inhibitor compound provided to the patient to the expected level of FADS1 inhibition.

[0664] Example 96 provided herein is the method of Example 95, further comprising determining an adjusted dose of the FADS1 inhibitor for the subject based on a comparison of the measured FADS1 inhibition level to the expected FADS1 inhibition level.

[0665] Example 97 provided herein is the method of Example 96, further comprising administering the adjusted dose of the FADS1 inhibitor to the subject.

[0666] Example 98 provided herein is a compound represented by the following structure:

[0667]

[0668] in

[0669] Each "*" symbol indicates an isotope that can be enriched 13 The location of C; and

[0670] At least one "*" position is isotopically enriched 13 C.

[0671] Example 99 provided herein is a compound represented by the following structure:

[0672]

[0673] in

[0674] Each "*" symbol indicates an isotope that can be enriched 13 The location of C; and

[0675] At least one "*" position is isotopically enriched 13 C.

[0676] Example 100 provided herein is a compound represented by the following structure:

[0677]

[0678] in

[0679] X is a halogen;

[0680] Each "*" symbol indicates an isotope that can be enriched 13 The location of C; and

[0681] At least one "*" position is isotopically enriched 13 C.

[0682] Embodiment 101 provided herein is the compound of any one of Embodiments 98 to 100, wherein at least two "*" positions are isotopically enriched 13 C.

[0683] Example 102 provided herein is the compound of any one of Examples 98 to 100, wherein at least three "*" positions are isotopically enriched 13 C.

[0684] Example 103 provided herein is the compound of any one of Examples 98 to 100, wherein at least four "*" positions are isotopically enriched 13 C.

[0685] Example 104 provided herein is the compound of any one of Examples 98 to 100, wherein all five "*" positions are isotopically enriched 13 C.

[0686] Embodiment 105 provided herein is the compound of any one of embodiments 98 to 104, wherein the enrichment factor for each position that is isotopically enriched is equal to or at least 100. For example, the isotopic enrichment factor at the labeled position is equal to or greater than 100.

[0687] Embodiment 106 provided herein is the compound of any one of embodiments 98 to 104, wherein the enrichment factor for each position that is isotopically enriched is equal to or at least 500. For example, the isotopic enrichment factor at the labeled position is equal to or greater than 500.

[0688] Embodiment 107 provided herein is the compound of any one of embodiments 98 to 104, wherein the enrichment factor for each position that is isotopically enriched is equal to or at least 1000. For example, the isotopic enrichment factor at the labeled position is equal to or greater than 1000.

[0689] Method for preparing labeled DGLA

[0690] Several embodiments disclosed herein provide methods for preparing labeled DGLA. In several embodiments, DGLA is prepared as described elsewhere herein. 13 C mark. Found that too little 13 C-atom labeled DGLA does not provide sufficient signal to measure the ratio of labeled DGLA to labeled AA (e.g., for pharmacodynamic analysis). Advantageously, it was found that having multiple (e.g., two, three, four, or five) 13 C-labeled DGLA provides sufficient signal to allow calculation of inhibitory concentration values (e.g., as a measure of the direct response distinct from endogenous DGLA and AA levels). 13 C-labeled DGLA provided sufficient signal to allow calculation of inhibitory concentration values. 13 C labeling (e.g., dihomo-γ-linoleic acid 1,2,3,4,5- 13 C) DGLA.

[0691] The following scheme (Scheme 1) provides an exemplary route for the synthesis of DGLA, although other routes will be readily apparent based on this disclosure:

[0692] Solution 1.

[0693]

[0694] Each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13C. In several embodiments, y is equal to or at least 1, 2, 3 or 4. For example, when y is 2, the two "*" symbols represent the enriched isotope. 13 The position of C (which can be expressed as [ 13 C2]). In some embodiments, at least two "*" symbols represent enriched isotopes. 13 In some embodiments, at least three "*" symbols indicate enriched isotopes. 13 In some embodiments, at least four "*" symbols represent enriched isotopes. 13 C position. In several embodiments, for the structure [ 13 C w ]-14 and [ 13 C w ]-15, each of the five "*" symbols represents an enriched isotope 13 The position of C.

[0695] The following scheme (Scheme 2) provides another exemplary route for the synthesis of DGLA, where each "*" is isotopically enriched. 13 C:

[0696] Option 2.

[0697]

[0698] In several embodiments, manufacturing 13 C w -DGLA (shown in the following structure):

[0699]

[0700] Including compounds

[0701]

[0702] Reacts with KC*N, each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13 C; and wherein X is a suitable leaving group. In several embodiments, X is halogen. In several embodiments, X is Br. In several embodiments, 13 C w -At least two "*" symbols on DGLA indicate enriched isotopes 13 C position. In several embodiments, 13 C w -At least three "*" symbols on DGLA indicate enriched isotopes 13 C position. In several embodiments, 13 C w-At least four "*" symbols on DGLA indicate enriched isotopes 13 C position. In several embodiments, 13 C w -All five "*" symbols on DGLA indicate enriched isotopes 13 The position of C.

[0703] Labeled 1,4-butanediol (e.g., 1,4-butanediol- 13 C4) is commercially available. It can be converted into compound [ 13 C4]-8:

[0704] Option 3.

[0705]

[0706] Example 108 of the present invention provides a method for producing a compound represented by the following structure:

[0707]

[0708] The method comprises making the following compound

[0709]

[0710] Reacts with KC*N;

[0711] in

[0712] Each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13 C; and

[0713] Wherein X is a suitable leaving group. In several embodiments, equal to or at least two "*" symbols indicate isotope enrichment 13 In some embodiments, equal to or at least three "*" symbols indicate enriched isotopes 13 In some embodiments, equal to or at least four "*" symbols indicate enriched isotopes 13 In several examples, each of the five "*" symbols represents an enriched isotope. 13 The position of C.

[0714] Embodiment 109 provided herein is the method of Embodiment 108, wherein X is halogen.

[0715] Presented herein as Embodiment 110 is the method of Embodiment 109, wherein X is Br.

[0716] Formulation and route of administration

[0717] Although it is possible to administer the compounds disclosed herein alone for the purposes described, the compounds typically administered will be present as active ingredients in pharmaceutical compositions. Thus, in one embodiment, provided herein are pharmaceutical compositions comprising the compounds disclosed herein in combination with one or more pharmaceutically acceptable excipients (e.g., diluents, carriers, adjuvants, etc.) and other active ingredients (if desired). In several embodiments, the pharmaceutical compositions comprise a therapeutically effective amount of a compound disclosed herein (e.g., a FADS1 inhibitor compound).

[0718] One or more compounds disclosed herein can be administered by any suitable route, in the form of a pharmaceutical composition suitable for such route, and in a dose effective for the intended treatment. The compounds and compositions presented herein can be administered, for example, orally, mucosally, topically, transdermally, rectally, pulmonary, parenterally, intranasally, intravascularly, intravenously, intraarterially, intraperitoneally, intrathecally, subcutaneously, sublingually, intramuscularly, intrasternally, intravaginally, or by infusion techniques in dosage unit formulations containing conventional pharmaceutically acceptable excipients.

[0719] The pharmaceutical composition can be in the form of, for example, tablets, chewable tablets, mini-tablets, caplets, pills, beads, hard capsules, soft capsules, gelatin capsules, granules, powders, lozenges, patches, creams, gels, sachets, microneedle arrays, syrups, flavored syrups, juices, drops, injectable solutions, emulsions, microemulsions, ointments, aerosols, aqueous suspensions, or oily suspensions. The pharmaceutical composition is typically prepared in dosage unit form containing a specific amount of the active ingredient.

[0720] Examples

[0721] Example A.1 provided herein is a method of identifying a subject having increased fatty acid desaturase 1 (FADS1) activity, the method comprising:

[0722] measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject;

[0723] wherein the one or more biological indicators comprise one or more of, consist of one or more of, or consist essentially of one or more of: a ratio of polyunsaturated fatty acids ("PUFAs"), a relative abundance of one or more cell types, a relative abundance of one or more differentially expressed genes ("DEGs") or gene signatures, and / or a relative abundance of one or more metabolites in the subject.

[0724] Example A.2 provided herein is a method of identifying a subject in need of treatment with a FADS1 inhibitor compound, the method comprising:

[0725] measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject;

[0726] wherein the one or more biological indicators comprise one or more of, consist of one or more of, or consist essentially of one or more of: a ratio of polyunsaturated fatty acids ("PUFAs"), a relative abundance of one or more cell types, a relative abundance of one or more differentially expressed genes ("DEGs") or gene signatures, and / or a relative abundance of one or more metabolites in the subject.

[0727] , Example A.3 provided herein is a method for reducing weight, reducing body mass index, treating obesity, treating metabolic disorders, treating cardiovascular disorders, treating diabetes, treating dyslipidemia and / or treating non-alcoholic steatohepatitis (NASH) in a subject, the method comprising:

[0728] measuring one or more biological indicators of a FADS1-mediated disease or disorder in the subject;

[0729] wherein the one or more biological indicators comprise one or more of, consist of one or more of, or consist essentially of one or more of: a ratio of polyunsaturated fatty acids ("PUFAs"), a relative abundance of one or more cell types, a relative abundance of one or more differentially expressed genes ("DEGs") or gene signatures, and / or a relative abundance of one or more metabolites in the subject.

[0730] Embodiment A.4 provided herein is the method of any one of Embodiments A.1 to A.3, further comprising administering to the subject a FADS1 inhibitor compound.

[0731] Embodiment A.5 provided herein is the method of any one of embodiments A.1 to A.4, wherein the ratio of PUFAs comprises or consists of an arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA) ratio.

[0732] Embodiment A.6 provided herein is the method of Embodiment A.5, wherein the AA to DGLA ratio is equal to or at least about 5:1.

[0733] Embodiment A.7 provided herein is the method of Embodiment A.5, wherein the AA to DGLA ratio is equal to or at least about 6:1.

[0734] Embodiment A.8 provided herein is the method of Embodiment A.5, wherein the AA to DGLA ratio is equal to or at least about 7:1.

[0735] Embodiment A.9 provided herein is a method of any one of embodiments A.1 to A.8, wherein the one or more cell types include one or more of adipocytes (Adipo), B cells (Bcell), endothelial cells (Endo), hepatocytes (Hep), Kupffer cells (Kupff), bone marrow cells (Myel), natural killer cells (NK), T cells (Tcell), or any combination of the foregoing.

[0736] Embodiment A.10 provided herein is the method of embodiment A.9, wherein the relative abundance of at least one cell type in the subject is increased.

[0737] Embodiment A.11 provided herein is the method of any of embodiments A.9 or A.10, wherein the relative abundance of at least one cell type in the subject is decreased.

[0738] Embodiment A.12 provided herein is the method of any one of embodiments A.1 to A.11, wherein the one or more metabolites comprise, consist of, or consist essentially of one or more of plasma cholesterol, free cholesterol, total cholesterol, cholesterol esters, malate, alpha-ketoglutarate, mannose, glucose, erythro-dihydrosphingosine, 5-O-methylsphingosine, threo-sphingosine, 1-hydroxy-2-amino-(cis,trans)-3,5-octadiene, 4-hydroxysphingosine, thromboxane B2, delta-12-prostaglandin D2, prostaglandin E 2. Prostaglandin D2, 12-hydroxyheptadecatrienoic acid, 14,15-dihydroxyeicosatrienoic acid, 11-hydroxyeicosatetraenoic acid, 13-hydroxyoctadecadienoic acid, arachidonic acid, docosahexaenoic acid, dihomo-gamma-linolenic acid, gamma-linolenic acid, docosapentaenoic acid, eicosapentaenoic acid, docosatatetraenoic acid, stearic acid, tryptophan, histidine, valine, threonine, cysteine, kynurenic acid, taurochenodeoxycholic acid, taurocholic acid, plasma triglycerides, plasmalogens, choline plasmalogens, inositol phospholipids, glycerol phosphate, phosphoric acid, lysophosphatidylcholine, lysophosphatidylethanolamine, and / or phosphatidylcholine.

[0739] Embodiment A.13 provided herein is the method of embodiment A.12, wherein the relative abundance of at least one metabolite in the subject is increased.

[0740] Embodiment A.14 provided herein is the method of embodiment A.12 or A.13, wherein the relative abundance of at least one metabolite in the subject is decreased.

[0741] Embodiment A.15 provided herein is the method of any one of embodiments A.1 to A.14, wherein the DEG comprises one or more of Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Hsd11b1, Akrld1, Aldh1a1, Mmp19, or a combination of any of the foregoing.

[0742] Embodiment A.16 provided herein is the method of any one of Embodiments A.1 to A.15, wherein the DEG comprises one or more of Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel1l3, Agap2, 4833411C07Rik, or a combination of any of the foregoing.

[0743] Embodiment A.17 provided herein is the method of any one of embodiments A.1 to A.16, wherein the DEG comprises one or more of: Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, or a combination of any of the foregoing.

[0744] Embodiment A.18 provided herein is the method of any one of Embodiments A.1 to A.17, wherein the DEG comprises one or more of: Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, or a combination of any of the foregoing.

[0745] Embodiment A.19 provided herein is the method of any one of Embodiments A.1 to A.18, wherein the DEG comprises one or more of TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Tagln2, or a combination of any of the foregoing.

[0746] Embodiment A.20 provided herein is the method of any one of Embodiments A.1 to A.19, wherein the DEG comprises one or more of Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, or a combination of any of the foregoing.

[0747] Embodiment A.21 provided herein is the method of any one of Embodiments A.1 to A.20, wherein the DEG comprises one or more of Cyp2b9, Fads1, Hao2, Cyp2b13, Cyp2a22, Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, or a combination of any of the foregoing.

[0748] Embodiment A.22 provided herein is the method of any one of Embodiments A.1 to A.21, wherein the DEG comprises one or more of Fitm1, Tmem86a, Agap2, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, or a combination of any of the foregoing.

[0749] Embodiment A.23 provided herein is the method of any one of Embodiments A.1 to A.22, wherein the DEG comprises one or more of: Ly6f, Foxil, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, or a combination of any of the foregoing.

[0750] Embodiment A.24 provided herein is the method of any one of Embodiments A.1 to A.23, wherein the DEG comprises Ppp2r5b.

[0751] Embodiment A.25 provided herein is the method of any one of Embodiments A.1 to A.24, wherein the DEG comprises one or more of: Slc6a7, Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Mfap2, or a combination of any of the foregoing.

[0752] Embodiment A.26 provided herein is the method of any one of Embodiments A.1 to A.25, wherein the DEG comprises one or more of Gm18537, Aacs, Pclo, Adrb3, Gm38394, AC154232.2, Cadps, Adgrb2, Gm45470, or a combination of any of the foregoing.

[0753] Embodiment A.27 provided herein is the method of any one of Embodiments A.1 to A.26, wherein the DEG comprises one or more of: Sdr9c7, Dsg1c, Slc17a1, Acnat2, Ces1c, Gss, Hsd11b1, or a combination of any of the foregoing.

[0754] Embodiment A.28 provided herein is the method of any one of Embodiments A.1 to A.27, wherein the DEG comprises one or more of 1810008I18Rik, Tlcd1, Snrk, Akrlc20, Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acss2, Acacb, Arhgap27, or a combination of any of the foregoing.

[0755] Embodiment A.29 provided herein is the method of any one of embodiments A.1 to A.28, wherein the DEG comprises one or more of Adam7, Rnase9, Wfdc8, Ighv9-2, DerI3, Acap1, Ccl19, Tcf7, Xkrx, Trim46, or a combination of any of the foregoing.

[0756] Embodiment A.30 provided herein is the method of any one of Embodiments A.1 to A.29, wherein the DEG comprises one or more of: Zfp369, Zfp871, Pcdhb21, Adrb3, Gm14288, Uprt, Atm, Dchs2, Cped1, Gm38357, or a combination of any of the foregoing.

[0757] Embodiment A.31 provided herein is the method of any one of Embodiments A.1 to A.30, wherein the DEG comprises one or more of: Cck, Ckap2, Gm4419, 1600015I10Rik, Sez6I2, Prnd, Gm16702, S100a8, or a combination of any of the foregoing.

[0758] Embodiment A.32 provided herein is the method of any one of Embodiments A.1 to A.31, wherein the DEG comprises one or more of: Pcdh12, Malat1, Kcnq1ot1, ArI4c, Gm38394, Gm42549, AC154232.2, Gm37310, Gm37776, Atp2a1, Ckm, Tnnt3, or a combination of any of the foregoing.

[0759] Embodiment A.33 provided herein is the method of any one of embodiments A.1 to A.32, wherein the DEG comprises one or more of Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Hsd11b1, Akrld1, Aldh1a1, Mmp19, or any combination of the foregoing Adipoq, Fabp4, Lep, Retn, Hoxc8, Hoxc9, Cebpa, Dgat1, Dgat2, Elovl3, Fads1, Fads2, Fas, Scd1, Srebf1, A drb3, Hilpda, Lipe, Mgll, Plin1, Plin4, Pnpla2, Pnpla3, Ldah, Cs, Gckr, Mel, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppara, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, Tm6sf2, Tnf, or a combination of any of the foregoing.

[0760] Embodiment A.34 provided herein is the method of any one of Embodiments A.1 to A.33, wherein the DEG comprises one or more DEGs as disclosed in any one or more of Figures 4a, 5a, 10a, and / or 13a.

[0761] Embodiment A.35 provided herein is the method of any one of embodiments A.1 to A.34, wherein the relative abundance of at least one DEG in the subject is upregulated.

[0762] Embodiment A.36 provided herein is the method of embodiment A.35, wherein the up-regulation factor of the at least one up-regulated DEG is equal to or greater than 2.

[0763] Embodiment A.37 provided herein is the method of any one of embodiments A.1 to A.36, wherein the relative abundance of at least one DEG in the subject is downregulated.

[0764] Embodiment A.38 provided herein is the method of embodiment A.37, wherein the down-regulation factor of the at least one down-regulated DEG is equal to or greater than 2.

[0765] Provided herein is embodiment A.39 which is the method of any one of embodiments A.1 to A.38, wherein the ratio of PUFAs comprises or consists of: a ratio of labeled DGLA to labeled AA;

[0766] wherein the DGLA to AA ratio is measured by administering a dose of labeled DGLA to the subject and then measuring the ratio of labeled AA to labeled DGLA.

[0767] Embodiment A.40 provided herein is the method of Embodiment A.39, wherein the labeled DGLA comprises isotopically enriched 13 C-labeled DGLA.

[0768] Embodiment A.41 provided herein is the method of Embodiment A.40, wherein the labeled DGLA comprises DGLA having equal or at least two positions of isotopically enriched 13 C atoms.

[0769] Embodiment A.42 provided herein is the method of Embodiment A.40, wherein the labeled DGLA comprises DGLA having equal to or at least three positions of isotopically enriched 13 C atoms.

[0770] Embodiment A.43 provided herein is the method of Embodiment A.40, wherein the labeled DGLA comprises DGLA having equal to or at least four positions of isotopically enriched 13 C atoms.

[0771] Embodiment A.44 provided herein is the method of Embodiment A.40, wherein the labeled DGLA comprises DGLA having equal to or at least five positions of isotopically enriched 13 C atoms.

[0772] Embodiment A.45 provided herein is the method of Embodiment A.40, wherein the labeled DGLA comprises DGLA uniformly enriched in isotopically 13 C atoms.

[0773] Embodiment A.46 provided herein is the method of any one of Embodiments A.39 to A.44, wherein the labeled DGLA comprises labeling at one or more or all of the regions shown below:

[0774]

[0775] Embodiment A.47 provided herein is the method of any one of Embodiments A.39 to A.46, wherein a dose of a FADS1 inhibitor compound is administered to the subject prior to, concurrently with, or after administration of the labeled dose of DGLA to the subject.

[0776] Example A.48 provided herein is the method of Example A.46, wherein the conversion of labeled DGLA to labeled AA is used to measure and / or calculate the level of inhibition of the FADS1 enzyme by the dose of the FADS1 inhibitor compound provided to the patient.

[0777] Embodiment A.49 provided herein is the method of Embodiment A.48 further comprising comparing the measured level of inhibition of the dose of the FADS1 inhibitor compound provided to the patient to the expected level of FADS1 inhibition.

[0778] Embodiment A.50 provided herein is the method of embodiment A.49, further comprising determining an adjusted dose of the FADS1 inhibitor for the subject based on a comparison of the measured FADS1 inhibition level to the expected FADS1 inhibition level.

[0779] Embodiment A.51 provided herein is the method of Embodiment A.50, further comprising administering to the subject the adjusted dose of the FADS1 inhibitor.

[0780] Example A.52 provided herein is a compound represented by the following structure:

[0781]

[0782] in

[0783] Each "*" symbol indicates a location that can be enriched with the isotope 13C; and

[0784] At least one "*" position is enriched in the isotope 13C.

[0785] Example A.53 provided herein is a compound represented by the following structure:

[0786]

[0787] in

[0788] Each "*" symbol indicates a location that can be enriched with the isotope 13C; and

[0789] At least one "*" position is enriched in the isotope 13C.

[0790] Example A.54 provided herein is a compound represented by the following structure:

[0791]

[0792] in

[0793] X is a halogen;

[0794] Each "*" symbol indicates a location that can be enriched with the isotope 13C; and

[0795] At least one "*" position is enriched in the isotope 13C.

[0796] Embodiment A.55 provided herein is the method of any one of Embodiments A.52 to A.54, wherein at least two of the "*" positions are isotopically enriched with 13C.

[0797] Embodiment A.56 provided herein is the method of any one of Embodiments A.52 to A.54, wherein at least three of the "*" positions are isotopically enriched with 13C.

[0798] Embodiment A.57 provided herein is the method of any one of Embodiments A.52 to A.54, wherein at least four of the "*" positions are isotopically enriched with 13C.

[0799] Embodiment A.58 provided herein is the method of any one of Embodiments A.52 to A.54, wherein at least five of the "*" positions are isotopically enriched with 13C.

[0800] Embodiment A.59 of the present invention provides a method for producing a compound represented by the following structure:

[0801]

[0802] The method comprises making the following compound

[0803]

[0804] Reacts with KC*N;

[0805] in

[0806] Each "*" symbol indicates a position that can be enriched with the isotope 13C and at least one "*" position is enriched with the isotope 13C; and

[0807] wherein X is a suitable leaving group.

[0808] Embodiment A.60 provided herein is the method of Embodiment A.59 wherein X is halogen.

[0809] Embodiment A.61 provided herein is the method of Embodiment A.60 wherein X is Br.

[0810] The following examples are given for the purpose of illustrating various embodiments of the present disclosure and are not intended to limit the present disclosure in any way. Those skilled in the art will readily appreciate that the present disclosure is well suited to achieving the ends and obtaining the objects and advantages mentioned, as well as those objects, ends and advantages inherent herein. Those skilled in the art will envision variations and other uses encompassed within the spirit of the present disclosure as defined by the scope of the claims.

[0811] Examples

[0812] This section provides specific examples of compounds having formula (I) and methods for their preparation.

[0813] Materials and Methods

[0814] Exemplary materials and methods used in the accompanying examples are provided below.

[0815] Reagents

[0816] List of chemical abbreviations

[0817] Ac Acetyl DCM dichloromethane DMSO dimethyl sulfoxide <![CDATA[Et2O]]> Diethyl ether EtOAc Ethyl acetate EtOH ethanol KHMDS Potassium hexamethyldisilazide NaHMDS Sodium hexamethylsilicon nitride <![CDATA[NaHCO3]]> Sodium bicarbonate <![CDATA[Na2SO4]]> sodium sulfate TsOH Toluenesulfonic acid TBAF Tetra-n-butylammonium fluoride TBDPS tert-Butyldiphenylsilyl TBDPSCl tert-Butyldiphenylchlorosilane THP Tetrahydropyran <![CDATA[TEA or Et3N]]> Triethylamine THF Tetrahydrofuran

[0818] FADS1 (NM_013402) and FADS2 (NM_004265) BacMam reagents were produced by the Biological Products Division of Amgen (Thousand Oaks, CA). 13 C1-C5] arachidonic acid ( 13 C5-AA), α-linolenic acid-d14 (ALA-d14), anandamide-d8 (AEA-d8), and polyunsaturated fatty acid standards were purchased from Cayman Chemical (Ann Arbor, Michigan). 13 C1-C5]8,11,14-eicosatrienoic acid ( 13 C5-DGLA) was custom synthesized by Curachem (Korea). 13 C 18 ]Linoleic acid( 13 C 18 -LA) was purchased from IsoSciences (Ambler, Pennsylvania).

[0819] Human plasma samples

[0820] All human samples were collected with site-specific Institutional Review Board approval, and appropriate informed consent was obtained in accordance with all applicable laws and regulations. In all cases, the materials obtained exceeded standard clinical practice and standard of care. Patient identity and protected health information / identifying information were redacted from tissue and clinical data prior to submission. Inclusion criteria required that subjects be free of cancer, cardiovascular disease, and autoimmune disorders.

[0821] Compound A FADS1 small molecule inhibitor

[0822] "Compound A" 8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one was synthesized according to the reaction procedures described in WO 2021 / 108404 A1. Compound A was dissolved in dimethyl sulfoxide (DMSO) for in vitro activity assays and formulated in 2% hydroxypropyl methylcellulose (HPMC) and 1% Tween 80 for mouse studies.

[0823] Mouse model and care

[0824] Mice were housed in an AAALAC internationally accredited facility. Experimental care was performed in accordance with the Guide for the Care and Use of Laboratory Animals, 8th edition. Mice were housed in individually ventilated cage (IVC) systems (Innorack, Innovive or Greenline, Tecniplast) and placed on irradiated corncob bedding (Envigo Teklad 7097). The lighting in the animal housing room was maintained at a 12:12 hour light: dark cycle, and the ambient temperature and humidity ranged from 68°F to 79°F and 30% to 70%, respectively. Animals had ad libitum access to food and reverse osmosis (RO) chlorinated water (2 to 3 ppm) via an automatic watering system as directed. Male AGN-rFADS1KO knockout (Fads1 KO; Fads1 Gt(IST11525H2)Tigm , Gene Trap IST11525H2, Texas A&M Institute for Genomic Medicine) and WT C57BL / 6 mice were maintained at Charles River Laboratories (Hollister, CA) or Jackson Laboratory (Bar Harbor, ME). Only male mice were used in the studies to avoid confounding factors of the female estrous cycle. Mice were fed a standard normal diet provided by Charles River (5066; LabDiet, Inc.; St. Louis, MA) until 5–8 weeks of age. After arrival and acclimation for 1–2 weeks, DIO mice were placed on a HFD (60 kcal% fat, D12492, Research Diets, Brunswick, NJ) for 12 weeks or until the indicated time. Lean controls were fed a standard normal diet (Envigo Teklad Universal Soy Protein-Free Extruded Rodent Diet 2020X, Indianapolis, IN).

[0825] Phenotypic study of Fads1 KO mice

[0826] Fads1 KO and WT mice were fed an HFD for 12 weeks. Weekly body weights and weekly fat and lean body composition measurements were recorded throughout the study (EchoMRI, Houston, TX). Food intake was measured at 7 weeks on the HFD. Intraperitoneal (IP) GTTs were performed using 2 g / kg glucose after a 12-hour fast at 7 weeks on the HFD. Four-hour fasted retroorbital blood was collected on the day the mice switched to the HFD and when they received the diet for 6 and 12 weeks. At the end of the 12-week study, mice were euthanized by conscious decapitation and plasma, liver, EPI, and ING WAT tissues were collected.

[0827] Long-term FADS1 inhibition in DIO mice

[0828] WT DIO mice were randomized and administered vehicle (2% HPMC, 1% Tween 80) or a dose of Compound A (10 mg / kg or 30 mg / kg) daily by PO gavage for 54 days. Body weight was measured daily and used for dosing calculations. 4-hour fasting retroorbital blood samples were collected at baseline and on day 46 for analysis of insulin, cholesterol, and triglycerides. Average food consumption over 3 days was measured on days 39-41. All animals received 0.1 ml of 2 mM Tris(R) by IP injection 23.5 hours after the last PO gavage. 13 C5-DGLA and various treatments were performed to allow for a 30-min exposure before necropsy to determine residual in vivo FADS1 activity.

[0829] Indirect calorimetry studies

[0830] Twenty-four Fads1 KO and 24 WT mice were entered into a Comprehensive Laboratory Animal Monitoring System (CLAMS; Columbus Instruments, Columbia, OH – Oxymax Model 2018, 0233-004M, Oxymax for Windows v5.53 software, hardware configuration 190395) before the start of the HFD feeding phase and again at 6 and 12 weeks of HFD. After 12 weeks of HFD, WT animals were randomly divided into two groups within the system (96 hours after baseline measurements) based on body weight, oxygen consumption, carbon dioxide production, age, and RER: vehicle (2% HPMC, 1% Tween 80) or 30 mg / kg of Compound A. Body weight was measured daily, and food consumption was measured every two weeks. Animals were re-entered into the CLAMS on days 17-21 and 38-42. Oxygen consumption (VO2), carbon dioxide production (VCO2), RER, and energy expenditure (HEAT) were measured during this period. For simplicity, the data are shown with lights on (light) and lights off (dark) cycles (12 hours on and 12 hours off, respectively).

[0831] Plasmid lipid, glucose, and insulin measurements

[0832] Mice were bled from the retro-orbital sinus under conscious awareness, and blood was collected into EDTA plasma tubes. Plasma lipids were measured using an Olympus AU400e chemistry analyzer (Olympus America, Central Valley, Pennsylvania). Plasma insulin was measured using an ALPCO mouse high-range insulin ELISA (Alpco, Salem, New Hampshire). Blood glucose was measured using an AlphaTRAK blood glucose meter (Zoetis, Parsippany, New Jersey).

[0833] RNA sequencing analysis

[0834] RNA was extracted using the Qiagen RNeasy 96 Universal Kit (QIAGEN, Germantown, MD). The concentration and quality of the isolated RNA samples were assessed and determined on an Agilent 4200 TapeStation system with an RNA integrity value greater than 9.1. Adipose or liver RNA (250 ng) was used to prepare cDNA libraries using a protocol modified from the Illumina TruSeq Stranded mRNA Kit (Illumina, San Diego, CA). Briefly, the polyA + After RNA selection, fragmentation, and priming, the fragmented RNA is TM II (Thermo Fisher Scientific-Invitrogen) TM , Waltham, MA) and RNase-out (Thermo Fisher Scientific-Invitrogen TM , Waltham, Massachusetts) were transcribed into cDNA in a reverse transcription reaction. The products of the first-strand cDNA synthesis reaction were then converted to double-stranded cDNA and subjected to end-repair, A-tailing, and adapter ligation according to commercial specifications. The constructed libraries were amplified and barcoded using a PCR program: a denaturation step of 98°C for 30 seconds, 15 cycles of 95°C for 10 seconds, 60°C for 30 seconds, and 72°C for 30 seconds, and an extension cycle at 72°C for 5 minutes, followed by a hold step at 4°C. Libraries were sequenced on an Illumina HiSeq4000 with read lengths of 100 or 150 nt and a minimum depth of 30 million paired-end reads.

[0835] RNA-Seq sequencing reads from mouse samples were aligned and processed using the OSA aligner, which was implemented in the OmicSoft array suite (QIAGEN OmicSoft, Cary, NC) based on the mouse genome version GRCm38 and the gene model GENCODE v19. Read count quantification was performed at the gene level based on the OmicSoft implementation of RSEM. Downstream analysis using read count output was performed according to the standard RNA-seq workflow of the DESeq2 R / Bioconductor package. Briefly, the raw read counts were transformed and normalized using the variance stabilizing transformation (VST) method. Principal component analysis (PCA) was performed using the top 1000 variable genes using the plotPCA function. Differential expression analysis based on a negative binomial distribution was performed using the DESeq function.

[0836] Gene expression is expressed as normalized fragments per kilobase per million reads (FPKM). FPKM values were further normalized by refining the commonly used upper quartile method, which sets FPKM to 10 at the 70th percentile. Lowly expressed genes with fewer than five samples with an FPKM ≥ 1 were removed. Genes with a Benjamini-Hochberg (BH) adjusted P value < 0.01 and a fold change ≥ 2 or ≤ 0.5 were selected as DEGs. Volcano plots of the differential expression analysis results were generated using the EnhancedVolcano R package (github.com / kevinblighe / EnhancedVolcano).

[0837] Selected DEGs were annotated by IPA (Qiagen, Redwood City, CA) on canonical pathways and toxicity lists. Gene expression enrichment in Gene Ontology (GO) and KEGG was analyzed using the Cluster Profiler R package with default parameters.

[0838] Deconvolution analysis was performed using MuSiC software v0.2.0 to estimate relative cell type abundance in each mouse bulk tissue sample. The Tabula Muris single-cell RNA-seq dataset was used as a reference for the MuSiC algorithm. Separate analyses were performed for each tissue type, including epithelial and ingrained water area (WAT) and liver.

[0839] Plasma PUFA measurement

[0840] 10 μl of plasma or PUFA standard was diluted in a surrogate matrix (60 g / l bovine serum albumin in Dulbecco's phosphate-buffered saline [DPBS]) and mixed with 10 μl of ALA-d as an internal standard. 14Mix in a 96-well plate. 100 μl of 2N NaOH was added to the mixture for subsequent saponification at 65°C for 1 hour. The mixture was then acidified with 50 μl of formic acid and subsequently extracted with 500 μl of hexane twice. The organic extracts from the two extractions were combined and the solvent evaporated under nitrogen. The extract was then resuspended in 250 μl of 90% methanol and analyzed by liquid chromatography tandem mass spectrometry (LC-MS / MS) for the following PUFAs: LA, GLA, DGLA and AA, with ALA-d 14 PUFA peak areas were quantified using Analyst software (SCIEX, Framingham, MA).

[0841] Tissue protein extraction and biomarker analysis of tissue and plasma samples

[0842] According to the manufacturer's instructions, frozen liver and adipose tissue samples were ground into powder using a stainless steel Bessman tissue grinder (Spectrum Laboratory Products, Gardner, California), and the resulting tissue powder was stored in -80°C cryovials until extraction. A portion of each powdered tissue sample was extracted using an internal protein extraction buffer (50mM Tris buffer, pH 7.4, 0.1M NaCl, and 0.1% Triton X-100) containing a protease inhibitor cocktail (Millipore Sigma, Burlington, Massachusetts). The sample was homogenized at 6500rpm for 30 seconds using MagNALyser (Roche Diagnostics, Indianapolis, Indiana), placed at 4°C for 30 minutes, and then spun at 10,000rpm for 10 minutes in a refrigerated microcentrifuge. The resulting supernatant was taken out to clean a 1.5ml microcentrifuge tube and spun at 14,000rpm for 15 minutes again. The final supernatant was aliquoted into labeled cluster tubes and the protein concentration was determined using a standard BCA protein assay (Thermo Fisher Scientific, Waltham, MA) before storage at -80°C.

[0843] The levels of various analytes were quantified using separate aliquots of tissue protein extracts and final plasma samples. Multiple adipokines and hormones (MCP-1, leptin, insulin, and PAI-1) were assessed using multiple mouse-specific Luminex assays, while adiponectin was assessed using a single mouse-specific Luminex assay (Millipore Sigma, Burlington, Massachusetts). All commercial assays were performed according to the manufacturer's instructions. Data were obtained on a FlexMap 3D (Luminex Corporation, Austin, Texas). Tissue protein expression levels were normalized for every milligram of total protein.

[0844] Plasma metabolome analysis

[0845] Metanomics Health (Germany) processed and analyzed plasma samples from male Fads1 KO mice (n=4) and their WT (n=6) littermates fed a HFD for 11 weeks for metabolomic analysis using the MxP Broad Spectrum and MxP Eicosanoid platforms.

[0846] Hepatic triglyceride measurement

[0847] Liver total lipids were homogenized and extracted with a chloroform-methanol solution at a ratio of 2:1. The extracted lipids were dried under nitrogen and suspended in a 90% isopropanol, 10% Triton X-100 solution. Liver triglyceride levels (mg / g tissue) were determined using total triglyceride measurement kits from Wako Diagnostics (Richmond, VA) and Sigma-Aldrich (Burlington, MA).

[0848] Tissue endocannabinoid measurements

[0849] Approximately 200 mg of tissue per sample was used for endocannabinoid extraction. 10 μl of ice-cold 100 mM butylated hydroxytoluene (BHT; in methanol) and 0.5 ml of 100 nM AEA-d8 (in water) were added to each frozen tissue sample. The tissue sample solution containing BHT was immediately homogenized at 30 Hz for 3 minutes using a TissueLyser (Qiagen, Germantown, Maryland), followed by two consecutive 1.25 ml ice-cold ethyl acetate:hexane (9:1) extractions. The combined extracts were dried under nitrogen until completely dry. The dried samples were redissolved in 80% acetonitrile and subjected to LC / MS analysis. 10 μl of sample was injected into a Kinetex C18 2.1 x 100 mm, 2.6 μm column (Phenomenex, Torrance, CA). Mobile phase A was 0.2% acetic acid in water and mobile phase B was 0.1% formic acid in acetonitrile. The LC gradient was a 7-minute method with a flow rate of 0.2 ml / min and consisted of an isocratic period of 0 to 1 minute at 75% B, followed by a gradient of 75% to 100% from 1 to 4 minutes and 100% to 75% B from 4 to 7 minutes. Analyte peak areas were quantified using Analyst software (SCIEX, Framingham, MA).

[0850] 13 C5-DGLA([ 13 Preparation of C5]-15)

[0851] [ 13 Synthesis of C4]-2

[0852]

[0853] NaH (2.20 g, 55.0 mmol) was added to a 500 mL flask. The NaH was washed with hexane (decanted twice), and then THF (50 mL) was added thereto. The resulting suspension was cooled to 0°C. [ 13 C4]-1 (4.70g, 50.0mmol) in THF (15mL) solution, and the reaction mixture was warmed to room temperature. After vigorous stirring at room temperature for 2 hours, the reaction mixture was cooled to 0 ° C again, and TBDPS-Cl (13.7g, 50.0mmol) was added dropwise. The temperature was then raised to room temperature and stirred for 1 hour. The reaction mixture was quenched with 10% K2CO3 aqueous solution (about 30mL), extracted with Et2O, dried over Na2SO4, filtered and evaporated. The product was purified by SiO2 column chromatography (EtOAc: hexane = 1: 4) to give [ 13 C4]-2 (16.1 g, 97%). 1H NMR(chloroform-d)δ:7.67(4H,br m),7.4(6H,br m),4.12(1H,br s),3.85(2H,br d),3.49(2H,br d),1.81(2H,br m),1.50(2H,br m),1.06(9H,s).

[0854] [ 13 Synthesis of C4]-3

[0855]

[0856] To a 500 mL flask purged with nitrogen was added a solution of oxalyl chloride (5.47 mL, 62.9 mmol) in DCM (15 mL). The flask was cooled to -78°C. A solution of DMSO (4.47 mL, 62.9 mmol) in DCM (20 mL) was added dropwise and the resulting mixture was stirred at -78°C for another 30 minutes. Then [ 13 C4]-2 (16.1 g, 48.4 mmol) in DCM (50 mL) was added. The resulting mixture was stirred at -78 ° C for another 30 minutes. At this time, TEA (17.5 mL, 126 mmol) was added dropwise and the reaction mixture was allowed to reach room temperature. After stirring overnight at room temperature, the reaction mixture was quenched by adding water (30 mL), extracted with DCM, dried over Na2SO4 and filtered. The product was purified by SiO2 column chromatography (5% EtOAc in hexane) to give [ 13 C4]-3 (11.2 g, 70%). 1 H NMR(chloroform-d)δ:9.75(1H,dd),7.67(4H,br m),7.4(6H,br m),3.86(1H,br m),3.50(1H,br m),2.70(1H,br m),2.38(1H,br m),2.04(1H,br m),1.72(1H,br m),1.06(s,9H).

[0857] [ 13 Synthesis of C4]-5.1

[0858]

[0859] A solution of phosphonium 4.1 (17.5 g, 36.1 mmol) in THF (60 mL) was cooled to -78°C and KHMDS (37.7 mL, 37.7 mmol) was added dropwise. After stirring at -78°C for 10 minutes, the reaction mixture was allowed to warm to room temperature. After stirring for 30 minutes, the reaction mixture was cooled to -78°C again. 13C4]-3 (11.2 g, 33.9 mmol) was dissolved in THF (20 mL) and the reaction mixture was allowed to warm to room temperature. After stirring overnight at room temperature, the reaction mixture was quenched by adding water (30 mL), stirred for 15 minutes, then extracted with Et2O, dried over Na2SO4, and filtered. SiO2 column chromatography (EtOAc: hexane = 1:9) gave [ 13 C4]-5.1 (14.4 g, 93%). 1 H NMR(chloroform-d)δ:7.66(4H,br m),7.40(6H,br m),6.62(1H,br m),5.40(2H,br m),5.15(1H,br m),4.58(br m),3.84(2H,br m),3.71(1H,br m),3.49(br m),3.38(br m),2.34(br m),1.98(br m),1.78-1.65(br m),1.60-1.21(br m),1.06(s,9H).

[0860] [ 13 Synthesis of C4]-6.1

[0861]

[0862] Place [ 13 C4]-5.1 (14.4 g, 31.4 mmol), 150 mL of EtOH, and 0.5 g of Pd / C. The reaction mixture was hydrogenated with 1 atm of H2 (balloon) at room temperature overnight. After filtration through Celite and evaporation, the crude product was used in the next step without additional purification. 13 C4]-6.1: 13.6 g (94%). 1 H NMR(chloroform-d)δ:7.66(6H,br m),7.38(8H,br m),4.57(br m),3.84(2H,br m),3.72(1H,br m),3.48(br m),3.37(br m),1.90-1.21(br m),1.06(s,9H).

[0863] [ 13 Synthesis of C4]-7

[0864]

[0865] Towards coarse 13C4]-6.1 (13.6 g, 29.6 mmol) was dissolved in EtOH (200 mL) and TsOH (1.23 g, 6.47 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. Then, a saturated aqueous solution of NaHCO3 (50 mL) was added. The resulting mixture was extracted with Et2O, dried over Na2SO4, and filtered. SiO2 column chromatography (EtOAc: hexane = 1:9) gave [ 13 C4]-7 (6.53 g, 59%). 1 H NMR (chloroform-d) δ: 7.66 (4H, br m), 7.38 (6H, br m), 3.83 (1H, br m), 3.63 (2H, br m), 3.48 (1H, br m), 1.7-1.11 (11H, br m), 1.06 (s, 9H).

[0866] [ 13 Synthesis of C4]-8

[0867]

[0868] Into a 500 mL flask purged with nitrogen was placed a solution of oxalyl chloride (1.97 mL, 22.7 mmol) in DCM (15 mL). The flask was cooled to -78°C. A solution of DMSO (1.61 mL, 22.7 mmol) in DCM (10 mL) was added dropwise and the resulting mixture was stirred at -78°C for another 30 minutes. Then, [ 13 C4]-7 (6.53 g, 17.4 mmol) was dissolved in DCM (50 mL) and the resulting mixture was stirred at -78 ° C for an additional 30 minutes. Finally, TEA (6.31 mL, 45.5 mmol) was added dropwise and the reaction mixture was allowed to reach room temperature. After stirring at room temperature for 5 hours, the reaction mixture was quenched by adding water (30 mL), extracted with DCM, dried over Na2SO4 and filtered. SiO2 column chromatography (EtOAc: hexane = 1:9) gave [ 13 C4]-8 (5.73 g, 88%). 1 HNMR(chloroform-d)δ:9.75(1H,s),7.66(4H,br m),7.38(6H,br m),3.83(1H,br m),3.47(1H,brm),2.40(2H,br m),1.7-1.11(8H,br m),1.06(s,9H).

[0869] A-[ 13 Synthesis of C4]-11

[0870]

[0871] A solution of phosphonium 10 (9.99 g, 19.7 mmol) in THF (100 mL) was cooled to -78°C and NaHMDS (19.7 mL, 19.7 mmol) was added dropwise. After stirring at -78°C for 1.5 hours, the aldehyde [ 13 C4]-8 (5.73 g, 15.4 mmol) was dissolved in 30 mL of dry THF (30 mL) and the reaction mixture was allowed to reach room temperature. After stirring at room temperature for 2 hours, the reaction mixture was quenched by adding aqueous NH4Cl solution, extracted with Et2O, dried over Na2SO4, filtered and evaporated. SiO2 column chromatography (1% EtOAc in hexane) gave [ 13 C4]11 (4.63 g, 58%). 1 H NMR(chloroform-d)δ:7.66(4H,br m),7.38(6H,br m),5.36(5H,m),3.82(1H,br m),3.47(1H,br m),2.80(3H,br m),2.04(4H,br m),1.7-1.11(15H,br m),1.04(s,9H),0.88(4H,br m).

[0872] [ 13 Synthesis of C4]-12

[0873]

[0874] Towards[ 13 To a solution of C4]-11 (4.63 g, 8.89 mmol) in THF (50 mL) was added a solution of TBAF in THF (13.3 mL, 13.3 mmol). After stirring at room temperature for 3 hours, the reaction mixture was quenched with water, extracted with Et2O, dried over Na2SO4, filtered, and evaporated. SiO2 column chromatography (EtOAc: hexane = 1:9) gave [ 13 C4]-12 (2.36 g, 94%). 1 H NMR (chloroform-d) δ: 5.36 (5H, br m), 4.11 (1H, br s), 3.81 (1H, br m), 3.47 (1H, br m), 2.81 (3H, br m), 2.04 (4H, br m), 1.7-1.11 (16H, br m), 0.88 (3H, br m).

[0875] [ 13 Synthesis of C4]-13

[0876]

[0877] Will[ 13 A solution of C4]-12 (2.36 g, 8.35 mmol) in DCM (30 mL) was cooled to 0°C and CBr4 (3.73 g, 11.3 mmol) was added in one go. After 5 minutes at 0°C, a solution of PPh3 (2.84 g, 10.8 mmol) in DCM (10 mL) was added dropwise. The reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was poured into a mixture of EtOAc: hexane (300 mL, 1:9) and the resulting suspension was filtered through diatomaceous earth. SiO2 column chromatography (Hex) gave [ 13 C4]-13 (2.58 g, 89%). 1 HNMR(chloroform-d)δ:5.36(5H,br m),3.59(1H,br m),3.21(1H,br m),2.81(3H,br m),2.05(4H,br m),1.7-1.11(16H,br m),0.88(3H,br m).

[0878] [ 13 Synthesis of C5]-14

[0879]

[0880] [ 13 C4]-14 (2.58 g, 7.47 mmol) and [ 13 C] KCN (0.64 g, 9.68 mmol) in DMSO (20 mL) was stirred at 50°C overnight. The reaction mixture was then poured into water and extracted with EtOAc. The combined organic layers were washed with water, dried over Na2SO4, and filtered. SiO2 column chromatography (2% EtOAc in hexane) gave [ 13 C5]-14 (2.08 g, 95%). 1 HNMR(chloroform-d)δ:5.37(5H,br m),2.81(3H,br m),2.50(1H,br s),2.17-2.05(5H,br m),1.82(1H,br m),1.7-1.11(15H,br m),0.89(3H,br m).

[0881] [ 13 Synthesis of C5]-15

[0882]

[0883] Will[ 13C5]-14 (2.08 g, 7.11 mmol) and NaOH (1.14 g, 28.50 mmol) in a mixture of EtOH / water (30 mL, 4 / 1) were heated at 100°C for 20 hours. The reaction mixture was evaporated, dissolved in water, and 1N HCl aqueous solution was added to make an acidic solution (pH about 2). The product was extracted with DCM, dried over Na2SO4, and filtered. SiO2 column chromatography (EtOAc: hexane = 3:7) gave [ 13 C5]-15 (2.06 g, 93%). After two consecutive MPLC (medium pressure liquid chromatography) purifications (Wakogel, mobile phase: water / acetonitrile / acetic acid = 10 / 90 / 0.1, room temperature), 300 mg [ 13 C5]-15. 1 H NMR(chloroform-d)δ:5.97(br m,6H),2.81(br m,4H),2.51(br m,2H),2.18(br m,2H),2.05(br m,4H),1.80(br m,1H),1.54(br m,4H),1.49(br m,4H),1.30(br m,4H), 1.19(br m,2H), 0.89(brm,1H), m / z=310.39.

[0884] Overview

[0885] The following examples provide additional details about the experiments performed in this article. Briefly, to determine whether FADS1 activity is elevated with obesity, AA / DGLA was used as a surrogate marker to measure FADS1 activity in obese male humans and diet-induced obese (DIO) mice. Fads1 KO mice fed a HFD were then characterized. Indirect calorimetry, transcriptome, and metabolome analyses were used to understand the physiological and metabolic effects of FADS1 inhibition. In addition, pharmacological inhibition of FADS1 in DIO mice was analyzed using Compound A, a novel, selective, and potent small molecule inhibitor of FADS1. Overall, the following studies demonstrate that FADS1 inhibitors reproduce the phenotypes observed in FADS1-deficient humans and KO mice, and that such inhibitors have the potential to become drugs for the treatment of obesity and related comorbidities.

[0886] Example 1: FADS1 activity in obese humans and DIO mice

[0887] Levels of n-3 and n-6 fatty acids (members of the PUFA pathway, as shown in Figure 1(a)) were higher in lean [body mass index (BMI) < 25 kg / m 2 , n = 20], overweight (25 <BMI<30kg / m 2, n = 41) and obesity (BMI>30 kg / m 2 , n=8) were measured in the plasma of males (Tables 1.1 and 1.2).

[0888] Table 1.1. Summary of male blood donors

[0889] queue N Age (years) <![CDATA[BMI(Kg / m 2 )]]> thin 20 50.1±12.1 23.7±1.3 overweight 41 54.6±9.1 27.4±1.4 obesity 8 52.6±7.2 32.2±1.0

[0890] Data are expressed as mean ± SD.

[0891] Table 1.2. PUFA profile in human male plasma (n=8-41 / group)

[0892]

[0893]

[0894] Data are expressed as mean ± SD.

[0895] Circulating levels of AA and DGLA did not differ between lean, overweight, and obese male subjects (Table 1.2); however, the AA / DGLA ratio (a surrogate for n-6 FADS1 activity) was significantly increased in obese men relative to lean subjects (Figure 1(b)). Similarly, PUFAs measured in the plasma of lean and DIO C57Bl / 6 male mice (Table 1.3) demonstrated that n-6 FADS1 activity (AA / DGLA ratio) was significantly increased in DIO mice relative to lean mice (Table 1.3 and Figure 1(c)). No differences in the GLA / LA ratio (a surrogate for n-6 FADS2 (D6D) activity) were observed between obese, overweight, and lean human males (Figure 11(a)) or between DIO mice relative to lean mice (Figure 11(b)). The increased FADS1 activity observed in obese humans and DIO mice suggests that DIO mice are a suitable preclinical model to further understand the role of FADS1 at the molecular and pharmacological levels.

[0896] Table 1.3. PUFA profiles in plasma of lean and DIO male mice

[0897]

[0898]

[0899] Example 2: Fads1 KO mice are resistant to HFD-induced obesity due to increased energy expenditure

[0900] To investigate the effects of FADS1 inhibition on obesity, age-matched wild-type (WT) and Fads1 KO male littermates were fed an HFD for 12 weeks. Given previous reports that Fads1 KO mice rely on AA supplementation for survival, the viability of Fads1 KO mice on an HFD containing 0.064% AA (wt / wt) was tested. Notably, Fads1 KO mice exhibited complete viability when fed a 0.064% AA diet, with no observed health issues after 32 weeks. At the end of the feeding period, Fads1 KO mice weighed 12.8% less than their WT littermates (Figure 2(a)). Relative to WT mice, Fads1 KO mice exhibited a 15.8% reduction in fat mass and an 11.6% reduction in lean body mass (Figures 2(b)-(c)). Fads1 KO mice did not exhibit statistically significant increases in food intake compared to WT mice (Figure 2(d)). Compared with WT littermates, Fads1 KO mice had lower insulin levels (Figure 2(e)) and improved glucose tolerance during a glucose tolerance test (GTT) (Figure 2(f)). Fads1 KO mice also exhibited reduced plasma cholesterol levels before starting the HFD compared with WT littermates, and maintained lower cholesterol levels throughout the HFD feeding period (Figure 2(g)). Triglyceride levels were either not different (Figure 2(h)) or slightly lower (Figure 7(a)) between Fads1 KO and WT mice.

[0901] Because AA supplementation might affect the phenotype of Fads1 KO mice, WT and Fads1 KO mice were fed a HFD with or without 2% AA for 9 weeks. Supplementation of Fads1 KO with a HFD containing 2% AA completely reversed the anti-obesity phenotype, indicating that the metabolic phenotype of Fads1 deficiency is AA-dependent and that the lack of additional AA supplementation is crucial for the manifestation of the obesity-resistant phenotype of Fads1 KO mice on a HFD.

[0902] To investigate the physiological and metabolic effects of FADS1 deficiency, indirect calorimetry was performed on Fads1 KO and WT mice at the start of HFD feeding and after 6 and 12 weeks (Figure 3). Fads1 KO mice showed significantly reduced oxygen consumption and carbon dioxide production during the light cycle at 0 and 6 weeks (Figure 3(a)-(b)) and 0, 6, and 12 weeks (Figure 3(c)-(d)), respectively. After 12 weeks of HFD feeding, Fads1 KO mice showed significantly reduced RER during both the light and dark cycles (Figure 3(e)-(f)), indicating increased lipid oxidation in Fads1 KO mice relative to WT mice. Fads1 KO mice were more active during the dark cycle than WT mice at 12 weeks and exhibited higher dark cycle energy expenditure relative to WT mice after 6 and 12 weeks of HFD feeding (Figure 3(g)-(h)). Figures 3(i)-(l) provide graphs of energy expenditure (kcal / hour) during the light and dark cycles compared to body weight after 6 and 12 weeks of HFD feeding. Since there was no significant difference in food intake (Figure 2(d)), the increase in energy expenditure helps to explain the reason why Fads1KO animals are resistant to DIO.

[0903] Example 3: Changes in transcriptome and metabolome in liver, fat, and plasma of Fads1 KO mice

[0904] Liver, epithelial (EPI), and ing (ING) WAT were collected from HFD-fed male Fads1 KO and WT mice and subjected to RNA sequencing. First, RNA-Seq expression profiles of Fads1 KO mice compared to WT mice were characterized to define genotype-dependent expression differences and identify FADS1-related genes associated with obesity. Liver and WAT exhibited distinct genotype-specific transcriptional profiles (Figure 4(a), Figure 5(a)). The transcriptome changes in EPI WAT were greatest in Fads1 KO mice compared to WT mice, both in terms of the number of affected genes and tissue cellularity (Figures 4(a)-(b), Figure 5(a)). The cellularity of EPI WAT was altered in Fads1 KO mice, showing a greater endothelial cell population (54% vs. 38%) and fewer myeloid cells (13% vs. 27%) compared to WT mice (Figure 4(b)).

[0905] The gene expression changes observed in the livers of Fads1 KO mice are likely due to changes in hepatic gene expression rather than changes in cellular composition (Figures 4(b)-(c), Figure 5(a)). Compared with WT, there were 185 overexpressed and 126 underexpressed genes in the livers of Fads1 KO mice, some of which belong to metabolic pathways, including lipid metabolism (Dgat2, Elovl3, Fads2, Lipe), fatty acid metabolism (Acox1, Hadhb), and PPAR signaling (Cpt1a, Cpt1b, Cyp4a12b) (Figures 4(c), Figure 5(a)). For EPI WAT, the metabolic pathways with the greatest changes were regulatory lipolysis (Adrb3, Lipe, Mgll, Pnpla2, Pnpla3), lipid metabolism (Dgat1, Dgat2, Fas, Scd1, Srebf1), fatty acid metabolism (Acaa1a, Acads, Acox1, Ehhadh), and insulin signaling (Irs1, Irs2) in adipocytes (Figures 4(d), 5(a)). Significant changes in gene expression in the PI3K signaling, apoptosis, and inflammation (Adgre1, Agtr1a, CD14, CD68, Fdft1) pathways in EPI WAT of Fads1KO mice relative to WT mice were consistent with shifts in the expression of cellular subsets of myeloid and lymphoid lineage immune cells (Figures 4(b), 4(d), 5(a)).

[0906] Compared with WT mice, the gene expression of adiponectin (Adipoq) was significantly upregulated in EPI WAT of Fads1 KO mice (Figure 5(a)), which was consistent with the higher adiponectin protein concentration in EPI WAT (Figure 5(b)). The protein concentrations of pro-inflammatory markers such as PAI-1 and MCP-1 were reduced in EPI and ING WAT of Fads1 KO mice (Figures 5(a)-(b)).

[0907] Plasma PUFA measurements and metabolomic analysis of HFD-fed Fads1 KO mice were significantly different from those of WT mice (Figure 7(a)-(c), Table 3.1). Plasma FA concentrations decreased, with a downward trend across all four lipid subfractions: phospholipids, triglycerides, cholesterol esters, and free fatty acids (FFA) (Figure 6(a)), and the FA composition within each subfraction shifted (Figure 6(b)). Circulating palmitic acid and stearic acid levels were decreased in Fads1 KO mice compared with WT mice (Figure 7(a), Tables 3.1 and 3.2). AA and EPA levels, as well as downstream PUFA levels such as adrenic acid, docosapentaenoic acid, and docosahexaenoic acid, were significantly reduced in Fads1 KO mice (Figure 7, Table 3.1). The FADS1 substrate DGLA and the upstream PUFA GLA were significantly higher in Fads1 KO mice than in WT mice. All four eicosanoids measured in the cyclooxygenase (COX) branch (prostaglandin E2, prostaglandin D2, delta-12 prostaglandin D2, and thromboxane B2) were reduced in Fads1 KO mice compared to WT mice (Figure 7), whereas only a small number of eicosanoids measured in each of the less inflammatory lipoxygenase (two of eight) and cytochrome P450 (CYP) cyclooxygenase (two of five) branches were significantly reduced in Fads1 KO mice compared to WT mice. In addition, citric acid cycle intermediates (malate and α-ketoglutarate) were elevated in Fads1 KO mice compared to WT mice, a characteristic of increased oxidation, and plasma taurine bile acids (taurocholic acid and taurochenodeoxycholic acid) were decreased (Figure 7). Levels of choline plasmalogens, sphingolipids, and phospholipid metabolites were also reduced in Fads1 KO mice relative to WT mice (Figure 7).

[0908] Table 3.1. PUFA profiles in plasma of Fads1 KO and WT male mice

[0909]

[0910] Table 3.2. Fatty acid concentrations of WT and Fads1 KO plasma lipid subfractions

[0911]

[0912] It is noteworthy that, despite increased plasma levels of 3-hydroxyisobutyrate (a metabolite of valine degradation) and upregulation of BCAA catabolism genes (Bcat1, Bcat2, Bckdha, Bckdhb, Bckdk) in EPI WAT, plasma levels of branched-chain amino acids (BCAAs; valine, leucine, and isoleucine) continued to increase (Figures 5(a), 7). Increased plasma levels of histidine and tryptophan were also observed, which may be the result of reduced hepatic expression of rate-limiting enzymes in histidine degradation (Hal) and tryptophan degradation (Tdo2) (Figures 5(a), 7(a)). It is noteworthy that, although the increase in plasma histidine levels was accompanied by the expected increase in glutamate levels (Figure 7(a)), the increase in tryptophan levels was not accompanied by a decrease in downstream metabolites; instead, increases in kynurenic acid and xanthurenic acid were observed (Figure 7(a)). This may be due to the upregulation of tryptophan degradation genes such as Kyat3 and Kmo in WAT (Figure 5(a)). Figure 5(b) provides a comparison of plasma, EPI WAT, and ING WAT levels of adiponectin, leptin, PAI-1, and MCP-1 in WT versus Fadsl KO mice.

[0913] IPA analysis of the metabolome changes in Fads1 KO mice revealed a trend of downregulation of biological functions, such as inflammatory response, lipid accumulation, activation of myeloid cells and leukocytes, and production of reactive oxygen species (ROS) (Figure 7(b)). When comparing the tissue transcriptome and plasma metabolome of Fads1 KO mice relative to WT mice, similar alterations in these biological functions were also observed in the liver or EPI WAT of Fads1 KO mice (Figure 7(c)).

[0914] Circulating kynurenic acid was elevated in Fads1 KO mice ( Figure 7 ), suggesting that it could increase energy expenditure by activating GPR35 in adipose tissue, thereby stimulating lipid oxidation, thermogenesis, and reducing inflammation, thereby attenuating weight gain and improving insulin sensitivity. Figure 6(b) provides the fatty acid composition of total plasma and each of the four lipid subfractions in plasma from WT and Fads1 KO mice.

[0915] Example 4: FADS1 inhibition studies using small molecule FADS1 inhibitors

[0916] Based on the altered FADS1 activity in obese humans and mice and the protective effect against diet-induced obesity in Fads1 KO mice, FADS1 inhibition has become a research goal. A potent and orally bioavailable molecule, compound A (Figure 8(a)), was identified. This compound inhibited FADS1 activity in vitro with a half-maximal inhibitory concentration (IC) of 1.5 for human and mouse FADS1. 50 ) were 0.003 μM and 0.004 μM, respectively, and showed selectivity for FADS2 ( FIG8( b )).

[0917] In vitro FADS1 and FADS2 activity assays

[0918] HEK 293 cells were mixed with FADS1 BacMam or FADS2 BacMam reagent at a multiplicity of infection (MOI) of 25 in Dulbecco's modified Eagle's medium (DMEM), 10% fetal bovine serum (FBS), 1x penicillin-streptomycin-glutamine (PSG) and plated in 96-well poly-D-lysine-coated plates. The plates were incubated at 37°C, 5% CO2 for 24 hours. The cells were washed with DPBS and incubated with 90 ml of compound dilutions in DPBS at 37°C for 20 minutes under 5% CO2. 10 μL of substrate, 13 C5-DGLA (FADS1) or 13 C 18 -LA (FADS2) was added to the cells and incubated at 5% CO2, 37°C for 30 minutes (FADS1) or 45 minutes (FADS2). The cells were then washed with DPBS, 50 μL of 2N NaOH was added to each well, and the cells were incubated in a 65°C oven for 1 hour for saponification. After cooling to room temperature, 22 μL of formic acid was added, followed by 113 μL of acetonitrile, and 1.5 μM ALA-d was added. 14 The samples were then subjected to LC-MS / MS to detect the following FADS1-responsive analytes: 13 C5-DGLA and 13 The following analytes reacted with C5-AA or FADS2: 13 C 18 -LA, 13 C 18 -GLA, 13 C 18 -DGLA and 13 C 18 The sample was injected into a Poroshell 120EC-C18 3.0 x 50 mm, 1.9 μm id column (Agilent, Santa Clara, CA).

[0919] Analyst software (SCIEX, Framingham, MA) was used to quantify the analyte peak areas. The percentage of substrate to product conversion relative to the untreated control (percent control [POC]) was calculated. GraphPad Prism (GraphPad Software, San Diego, CA) was used to plot the nonlinear regression of POC versus compound concentration.

[0920] In vivo FADS1 activity measurement using liver and plasma

[0921] To measure the in vivo activity of FADS1 using the liver, dose titrated Compound A was orally administered to DIO C57B1 / 6 mice for 23.5 hours. The mice were then dosed by intraperitoneal injection of a solution containing 0.1 ml 13 C5-DGLA solution (2 mM in 18.42 mM Na2CO3 buffer). 13 Necropsy was performed 30 minutes after C5-DGLA administration. 13 The livers of DIO mice administered with C5-DGLA were used for pharmacodynamic (PD) analysis, and EDTA plasma was collected for compound exposure determination. For PD analysis, 150 mg of frozen liver tissue was mixed with ALA-D as an internal standard. 14 Premix. The liver sample was homogenized and extracted twice with a chloroform / methanol (2:1) solution. The organic phases of the two extractions were combined and evaporated to dryness under nitrogen. The liver extract was resuspended in 200 μl of 2N NaOH and saponified at 65°C for 1 hour. After cooling to room temperature, 100 μl of formic acid was added and the mixture was extracted twice continuously with a chloroform:methanol (2:1) solution. The combined organic phases were evaporated to dryness under nitrogen and resuspended in a solution containing acetonitrile and methanol in a ratio of 1:1. The extracts were then analyzed by LC / MS / MS for the following analytes: 13 C5-DGLA, 13 C5-AA, ALA-d 14 As internal standard. The sample (20 μl) was injected into a Poroshell 120EC-C18 3.0x 50mm, 1.9μm column (Agilent, Santa Clara, California). The analyte peak area was quantified using Analyst software (SCIEX, Framingham, Michigan). The residual in vivo FADS1 activity was calculated by normalizing the percent conversion of substrate to product to the vehicle group to determine the percent control (POC) and the percent inhibition relative to the control (vehicle).

[0922] To determine the extent of FADS1 inhibition, in vivo FADS1 activity (expressed as percent conversion) was calculated using the following formula:

[0923] Calculate conversion percentage = product / (substrate + product)*100

[0924] = 13 C5-AA / ( 13 C5-DGLA+ 13 C5-AA)*100.

[0925] The data were then normalized to the vehicle group to determine the percent inhibition relative to the control (vehicle) by the following equation:

[0926] Percent inhibition = (1 - (FADS1 activity per mouse / average FADS1 activity of vehicle group)) * 100

[0927] To calculate in vivo efficacy, a dose-response graph was constructed by plotting the plasma concentration of the test compound determined from the PK analysis on the abscissa relative to the percent control (POC, relative to vehicle) of FADS1 activity in each mouse. The data points were then fitted using a log(inhibitor) versus response nonlinear regression algorithm using GraphPad Prism (version 8.4.3, San Diego, CA) to calculate the in vivo IC 50 (total).

[0928] Free (unbound) IC 50 and IC 90 The values were calculated as follows: first, the plasma concentration of the test compound was multiplied by the fraction unbound (fu) value in mouse plasma determined in the plasma protein binding study to calculate the concentration of unbound compound in plasma. A dose-response curve was used to determine the unbound IC by plotting the unbound plasma concentration of the compound on the abscissa relative to the percent control (POC, relative to vehicle) of FADS1 activity in each mouse. 50 and IC 90 The data points were then fitted using a log(inhibitor) versus response nonlinear regression algorithm using GraphPad Prism (version 8.4.3, San Diego, CA) to calculate the in vivo IC 50 (free). IC in vivo 90 (Free) was calculated by using the generated curve to interpolate the concentration at which 90% of the in vivo FADS1 activity was inhibited and only 10% of the activity was observed.

[0929] In addition to using liver, plasma was also used to measure FADS1 activity in the presence of Compound BE. The above procedure was performed to measure the in vivo activity of FADS1 using liver. The following general procedure was used to measure FADS1 activity using plasma. Diet-induced obese (DIO) mice that had been fed a high-fat diet for 12 weeks were used to determine the pharmacokinetic (PK) and pharmacodynamic (PD) properties of the test compounds. To determine the target coverage 24 hours after administration of the test compound (e.g., Compound BE, as shown below), mice were given 0.1 ml of 13 C5-DGLA solution (2 mM in 18.42 mM Na2CO3 buffer). 13Necropsy was performed 30 minutes after C5-DGLA administration, and EDTA plasma was collected for PK and PD analysis.

[0930]

[0931] For PD analysis, 50 μl of plasma or standards (pooled untreated mouse plasma spiked with 4.57 nM to 10 μM of dapoxetine) were plated in a 96-well plate. 13 C5-DGLA and 13 C5-AA) were mixed with 10 μl of 90% methanol and 10 μl of 50 μg / mL ALA-D14 (as internal standard) (100 μM α-linolenic acid-d 14 (ALA-d 14 , Cayman Chemical Company)) mixed. 490μl chloroform: methanol (2: 1) was added to each well. The plate was placed on a shaker for 5 minutes, and then 66.5μl water was added. The plate was shaken for another 5 minutes and then centrifuged at 2000RPM for 5 minutes. The bottom organic layer was transferred to a clean 96-well plate. 333ul chloroform: methanol (2: 1) was added to the remaining liquid phase in the original plate. After shaking for 5 minutes, the plate was centrifuged at 2000RPM for 5 minutes. The bottom layer was collected and merged with the first bottom layer. The combined layer was evaporated to dryness under a constant nitrogen flow. After the plate was dried, 100μl of 2N NaOH was added to the mixture for subsequent saponification at 65°C for 1 hour. The sample was then cooled to room temperature, and the mixture was acidified with 50μl formic acid, followed by two consecutive chloroform: methanol (2: 1) extractions. 500μl chloroform: methanol (2: 1) was added and the mixture was thoroughly mixed by shaking. The plate was then centrifuged at 2,000rpm for 5 minutes. The bottom chloroform phase was transferred to a new 2 mL 96-well plate and the remaining aqueous layer was extracted with another 250 μl of chloroform: methanol (2: 1). The organic extracts were combined and the solvent was evaporated to dryness by placing the plate under nitrogen. The extract was then dissolved in 200 μl of acetonitrile / methanol (1: 1). The plate was placed on a shaker for 30 minutes to ensure that the extracted fatty acids were fully mixed and dissolved. The plate was then centrifuged at 2000 RPM for 5 minutes. 80 μl of sample was transferred to a new 96-well polypropylene plate and the following analytes were analyzed on LC-MS / MS: 13 C5-DGLA, 13 C5-AA, ALA-d 14as an internal standard. Briefly describe the LC-MS / MS method: 20 μl of sample was injected onto a Poroshell 120EC-C18 3.0 x 50 mm, 1.9 μm column. The mobile phases were 80% acetonitrile containing 5 mM ammonium acetate for mobile phase A and 99.5% acetonitrile containing 5 mM ammonium acetate for mobile phase B. The LC gradient was a 3-minute method with a flow rate of 0.4 mL / min and consisted of: 45% B from 0 to 0.5 minutes, followed by 45% B to 100% B from 0.5 to 2.0 minutes; the system then remained at 100% B from 2.0 to 2.5 minutes, returning to 45% B at 2.6 minutes and maintaining this level until the end of the method. Analyte peak areas were quantified using SCIEX Analyst software. To determine the extent of FADS1 inhibition, in vivo FADS1 activity, POC, percent inhibition, and in vivo IC50 values were calculated as described above for hepatic FADS1 activity measurements.

[0932] For each compound BE, 13 C5-DGLA to 13 The % conversion and % inhibition data for C5-AA were compiled using data from liver and plasma. Table 4.1 and Figure 17a provide the % conversion and % inhibition data for compound BE. 13 C5-DGLA to 13 The conversion % data of C5-AA were obtained using the C5-AA from liver and plasma, respectively. 13 C5-DGLA and 13 C5-AA measurement.

[0933] Table 4.1

[0934]

[0935] Table 4.2 and Figure 17b provide the % inhibition for compound BE, as measured using samples collected from liver and plasma, respectively. 13 C5-DGLA and 13 C5-AA measurement.

[0936] Table 4.2

[0937]

[0938]

[0939] Comparison of FADS1 activity in the liver and plasma of DIO mice 24 hours after oral administration of a single dose of each of Compounds B, C, D, and E revealed highly similar percent inhibition values between the two tissues, despite differences in the percent conversion detected, indicating that plasma FADS1 activity measurements reflect liver FADS1 activity. In other experiments (not shown), target coverage was measured (e.g., in time course studies) at 1, 2, 4, 6, 8, or 16 hours (or other time points) after administration of the test compound.

[0940] The results were obtained by measuring the effects of different doses of the corresponding compounds on the liver of DIO mice. 13 C5-DGLA and 13 C5-AA, calculated FADS1-unbound in vivo IC50 values for Compounds A, B, and C. The calculated IC50 values are shown in Table 4.3. An exemplary dose response curve for Compound A is provided in Figure 8(c).

[0941] Table 4.3

[0942] Compound <![CDATA[Free (unbound) in vivo IC 50 [μM]]]> Compound A 0.002 Compound B 0.004 Compound C 0.004

[0943] Using a similar procedure as described above for Compound AE, the plasma levels of Compound F (shown below) at different doses were measured. 13 C5-DGLA and 13 C5-AA, an additional in vivo FADS1 potency assay was performed. A dose response curve was generated, as shown in Figure 17c.

[0944]

[0945] Using plasma collected 13 C5-DGLA and 13 The inhibitory concentration values of compound F at C5-AA are as follows (Table 4.4):

[0946] Table 4.4

[0947]

[0948] Example 5: Compound A treatment reduces body weight and improves metabolic profile in DIO mice

[0949] The physiological and metabolic effects of long-term pharmacological inhibition of FADS1 by Compound A were evaluated in DIO mice. Long-term FADS1 inhibition for 54 days by oral administration of 10 mg / kg and 30 mg / kg of Compound A resulted in a 17.9% and 21.1% reduction in body weight, respectively, relative to vehicle-treated mice (Figure 8 (d)). FADS1 activity (measured after 54 days of treatment) was inhibited by 90.0% and 93.6% in the 10 mg / kg and 30 mg / kg Compound A treatment groups, respectively (Figure 8 (e)). No significant differences were observed in food intake (Figure 8 (f)). Compared to vehicle-treated mice, the plasma insulin levels of mice treated with 10 mg / kg and 30 mg / kg Compound A were significantly reduced by 74.9% and 84.7%, respectively (Figure 8 (g)). Plasma cholesterol showed a downward trend (Figure 8(h)), and a dose-dependent decrease in plasma triglycerides was observed, with levels reduced by 41.6% in mice treated with 30 mg / kg Compound A compared to vehicle-treated mice (Figure 8(i)). Similar changes in plasma FA and lipid subfraction composition were observed in Compound A-treated DIO mice (Figure 16(a), Tables 5.4, 5.5, and 5.6) as those observed in Fads1 KO mice (Figure 6(a), Tables 5.1, 5.2, and 5.3). Notably, the ratio of the two products of the FADS1 reaction, the EPA / AA ratio (a marker negatively correlated with inflammation and metabolic disease), increased in all four lipid subfractions (Table 5.5); a similar trend was observed in Fads1 KO mice (Table 5.2), indicating differences in the metabolism of EPA and AA.

[0950] Table 5.1. Plasma PUFA and metabolic profiles of Fads1 KO and WT male mice after 12 weeks of high-fat diet.

[0951]

[0952]

[0953] Polyunsaturated fatty acids (PUFA); wild type (WT); knockout (KO); fatty acids measured by LC / MS-MS; n = 8-9 / genotype; data presented as mean ± SD; ^unpaired t-test, *P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant.

[0954] Table 5.2. Fatty acid concentrations of WT and Fads1 KO plasma lipid subfractions

[0955]

[0956]

[0957] Wild type (WT); knockout (KO); n = 3 pooled samples / group; each sample was pooled from 2 mice fed a high-fat diet for 12 weeks; data are expressed as mean values in μg mL -1 ; Measured by GC (OmegaQuant, Sioux Falls, SD); Statistics: unpaired t-test for each subfraction relative to WT; *P<0.05, **P<0.01, ***P<0.001.

[0958] Table 5.3. Fatty acid composition of WT and Fads1 KO plasma lipid subfractions

[0959]

[0960]

[0961] Wild type (WT); knockout (KO); n = 3 pooled samples / group; each sample was pooled from 2 mice fed a high-fat diet for 12 weeks; data are expressed as the mean % composition of each lipid subfraction; measured by GC (OmegaQuant, Sioux Falls, SD); Statistics: unpaired t-test for each fraction relative to WT; *P < 0.05, **P < 0.01, ***P < 0.001

[0962] Table 5.4. Plasma PUFA and metabolic profiles of WT DIO mice treated with Compound A

[0963]

[0964]

[0965] Polyunsaturated fatty acids (PUFA); wild type (WT); diet-induced obesity (DIO); n = 3-10 / group, after 46 or 54 days of treatment; data are expressed as mean ± SD; fatty acids measured by LC-MS / MS; Statistics: one-way ANOVA with Dunnett's multiple comparison test relative to vehicle control; *P < 0.05, **P < 0.01, ***P < 0.001.

[0966] Table 5.5. Fatty acid concentrations in plasma lipid subfractions of Compound A-treated DIO mice

[0967]

[0968]

[0969] Wild type (WT); diet-induced obesity (DIO); WT vehicle-treated (WT); WT treated with 30 mg / kg (WT+A); n = 3 pooled samples / group; each sample was pooled from 2 WT DIO mice; data are expressed as mean values in μg mL -1 ; Measured by GC (OmegaQuant, Sioux Falls, SD); Statistics: Unpaired t-test relative to WT, vehicle-treated group for each subfraction; *P<0.05, **P<0.01, ***P<0.001.

[0970] Table 5.6. Fatty acid composition of plasma lipid subfractions in DIO mice treated with Compound A.

[0971]

[0972]

[0973] Wild type (WT); diet-induced obesity (DIO); WT vehicle-treated (WT); WT treated with 30 mg / kg (WT+A); n=3 pooled samples / group; each sample was pooled from 2 WT DIO mice; data are expressed as the mean % composition of each lipid subfraction; measured by GC (OmegaQuant, Sioux Falls, SD); Statistics: unpaired t-test relative to WT, vehicle-treated groups for each subfraction; *P<0.05, **P<0.01, ***P<0.001.

[0974] Indirect calorimetry of DIO mice treated with vehicle or 30 mg / kg Compound A for 42 days again significantly reduced body weight, as expected (Figure 9(a)); however, no differences were observed in energy expenditure, oxygen consumption, and carbon dioxide production (Figures 9(b)-9(f)). However, Compound A-treated mice had reduced light cycle RER in both measurement periods, 17-23 days and 38-42 days (Figures 9(g), 9(h)), indicating increased resting lipid oxidation. In addition, Compound A-treated mice were more active during the dark cycle than vehicle-treated mice (Figures 9(i)-9(j)).

[0975] Example 6: RNA-seq analysis in DIO mice treated with Compound A

[0976] Next, RNA-seq analysis was performed to compare the transcriptome changes caused by genetic deletion or pharmacological inhibition of FADS1. Comparison of transcriptome changes in DIO mice treated with 30 mg / kg compound A (WT+A) for 54 days and age-matched vehicle-treated DIO Fads1KO relative to vehicle-treated WT mice revealed similarities and differences in affected pathways (Figure 10). Figure 10 (a) provides a radar chart of commonly altered genes in liver, EPI WAT, and ING WAT of Fads1 KO mice in compound A-treated mice relative to WT vehicle-treated mice. Compared to WT, genetic inhibition of FADS1 activity and pharmacological inhibition of FADS1 activity by compound A resulted in a greater number of changes in the transcriptome in EPI WAT. The number of DEGs in the liver was moderate, and the number of DEGs in ING WAT was the least (Figure 13 (g)). Furthermore, genetic or pharmacological inhibition of FADS1 activity resulted in transcriptome changes in 122 common genes in liver, 872 in EPI WAT, and 40 genes in ING WAT (Figure 13(g)). Relative to WT mice, the cellular composition of liver and ING WAT in mice treated with Compound A was moderately affected. Following treatment with Compound A, the largest changes observed in the cellular composition of EPI WAT were in immune cell populations, with a decrease in T cells, B cells, and natural killer cells.

[0977] Ingenuity pathway analysis (IPA) of transcriptome changes achieved by genetic inhibition of FADS1 and pharmacological inhibition of FADS1 by compound A in DIO mice revealed some common pathways in EPI WAT: upregulation of the mTOR signaling pathway and downregulation of 23 pathways (Figure 10(c)). Fewer pathways were affected in the liver, and common affected pathways shared by Fads1KO and compound A-treated mice included upregulation of cholesterol biosynthesis and nicotine degradation pathways and reduction of CREB signaling and eNOS signaling pathways in neurons (Figure 10(d)). Unique pathways affected by genetic loss of Fads1 included methionine degradation in EPI WAT (Figure 10(c)) and three signaling pathways in the liver (GP6 signaling, xenobiotic metabolism CAR signaling, and PXR signaling) (Figure 10(d)). In EPI WAT, inhibition of FADS1 by compound A increased insulin receptor signaling (Figure 10(c)). In the liver of WT mice treated with compound A (Figure 10(d)), upregulation of calcium signaling, PPARα / RXRα activation, and xenobiotic metabolism AHR signaling pathways, as well as downregulation of acute phase response, AMPK signaling, and senescence pathways, were observed. Disease and biological function analysis of common DEGs in Fads1 KO mice and compound A-treated mice revealed common and differential biological functions affected in liver, EPI WAT, and ING WAT, including diabetes, diabetic nephropathy, and inflammation in absolute anatomical regions that were commonly altered in all three tissues.

[0978] Gene expression profiles of liver, EPI WAT, and ING WAT of Compound A-treated mice showed similar gene expression trends to those of Fads1KO mice: increased fatty acid oxidation-related genes (Cpt1a, Acox1, Acaa1a, Fgf21) in the liver; increased lipolysis genes (Lipe, Mgll, Pnpla2) and insulin signaling genes (Insr, Irs1, Slc2a4) in EPI WAT; and decreased inflammation-related genes (Ccl2, Cd14, Cd68) and upregulation of insulin signaling genes (Insr, Irs1) in ING WAT (Figures 13(a)-(c)). Upstream analysis of IPA revealed PPARA, PPARG, RXRA, and PPPARGC1A as top predictive upstream regulators activated in the liver (Figure 13(d)). Many inflammation-related upstream regulators, including IFNG, TCF3, and TNF, were predicted to be suppressed in EPI WAT of Fads1 KO and Compound A-treated mice ( FIG. 13( e ) ).

[0979] Compound A treatment reduced leptin levels in plasma and in EPI and ING WAT (Figure 13 (f)). This is different from Fads1 KO mice, which had slightly higher leptin levels in EPI WAT than WT (Figure 13 (f)). As observed in Fads1 KO mice, adiponectin levels were increased in EPI WAT of WT mice treated with Compound A. WT mice treated with Compound A showed a trend toward decreased PAI-1 and MCP-1 levels in EPI and ING WAT (Figures 13 (d)-(f)), which is consistent with the decreased expression of inflammation-related genes in the corresponding tissues (Figure 10, Figure 13 (c)).

[0980] discuss

[0981] Obesity is associated with chronic low-grade inflammation, and FADS1 synthesizes AA, a precursor to many proinflammatory eicosanoids. As demonstrated in the examples herein, FADS1 activity, as indicated by the plasma AA / DGLA ratio, was found to be elevated in obese male humans and male mice, indicating that FADS1 activity is elevated in the obese state (Figures 1(b)-(c)). Similarly, increased FADS1 (or D5D) activity was found in E3L mice fed a HFD. However, the data presented herein conflict with other human studies that report a lack of association or a negative correlation between FADS1 activity and obesity or insulin resistance. The differences in FADS1 activity found between different reports may reflect differences in the study population (i.e., disease state), the type of diet consumed by individuals, and other confounding factors. These differences may affect PUFA levels and the expression and activity of fatty acid desaturases. For example, when FADS2 activity is found to be positively correlated with BMI, a negative correlation between FADS1 activity and BMI is often detected. Since FADS2 is upstream of FADS1 and is the rate-limiting enzyme in this pathway, inhibiting FADS2 activity with the FADS2-specific inhibitor SC-26196 will increase the downstream AA / DGLA ratio ( Figure 11(c) ), thereby giving the illusion of increased FADS1 activity. Figure 12Inhibition of FADS2 activity by SC-26196 has been shown to affect not only the plasma GLA / LA ratio in DIO mice but also the plasma AA / DGLA ratio. Increased FADS2 activity could have the opposite effect. Therefore, it is hypothesized that in population-based studies, the true effect of FADS1 may be confounded by changes in FADS2 activity; obese subjects with elevated FADS2 activity may have a lower AA / DGLA ratio, leading to the apparent inverse association between FADS1 and obesity. Importantly, the FADS locus has been associated with body weight, type 2 diabetes, dyslipidemia, and other cardiometabolic traits in GWAS, suggesting an involvement of FADS1 in body weight and metabolic disease. The minor alleles of the FADS1 SNPs rs174556 and rs7115739, which are associated with reduced FADS1 activity, may be associated with lower body weight. Furthermore, carriers of the minor C allele of the FADS1 SNP rs174547 (possibly associated with reduced FADS1 activity) have lower odds of metabolic syndrome and greater waist circumference relative to carriers of the TT genotype. These together support the concept that FADS1 activity is positively correlated with obesity.

[0982] Here, ablation of FADS1 activity by genetic deletion or pharmacological inhibition reduced body weight and metabolic profiles with minimal effects on food intake. Using untargeted transcriptomic and metabolomic approaches, the data presented here reveal possible mechanisms of action, such as reduced inflammation and increased lipid oxidation through PPARα activation.

[0983] The improvements in metabolic parameters achieved by inhibiting FADS1 activity through genetic deletion or pharmacological inhibition may be mediated in part by suppressing inflammation by limiting the availability of AA (Tables 3.1 and 6.1) and AA-derived proinflammatory eicosanoids, particularly COX-derived eicosanoids (Figure 7(a)). COX has been implicated in the obesity-associated metabolic syndrome. Administration of indomethacin, a general COX inhibitor, to HFD-fed DIOC57BL / 6J mice reduced body weight and improved insulin sensitivity. Therefore, reduction in COX-derived eicosanoids may partially contribute to the anti-obesity effects of FADS1 inhibition.

[0984] The reduction in inflammation was particularly pronounced in adipose tissue of Fads1 KO and Compound A-treated mice. EPI and INGWAT analyses revealed downregulation of inflammatory pathways and biological functions, including monocyte / macrophage markers (Cd14 and Cd68) (Figures 4(d), 5(a), 7(c), 10(c), 13(c)-(d)), alterations in immune cell populations (Figures 4(b), 10(b)), and decreased PAI-1 and MCP-1 protein levels (Figures 5(b), 13(d)), suggesting that FADS1 inhibition reduces adipose macrophage infiltration and inflammation. This result is consistent with previous reports of downregulation of inflammatory genes in adipose tissue of FADS1-inhibited mice. Obesity leads to a dramatic increase in macrophages in human and mouse adipose tissue, resulting in increased production of inflammatory cytokines, which alter insulin signaling and contribute to insulin resistance in obesity; therefore, reducing adipose macrophage infiltration and inflammation by inhibiting FADS1 may improve insulin resistance in DIO mice. Insulin sensitivity was improved in Fads1 KO and Compound A-treated mice, as demonstrated by improved glucose tolerance, reduced blood glucose and insulin levels (Figures 2(e)-(f), 7, 8(g)), and upregulation of insulin signaling genes (Insr, Irs1, Slc2a4) in EPI and ING WAT (Figures 4(d), 10(c), 5(a)). Confirming data were also reported in humans; carriers of the minor allele genotype (CC) of the FADS1 SNP rs174550 were found to be protected from LA-induced adipose tissue inflammation and had improved insulin sensitivity through lower FADS1 activity. Together, these data suggest that FADS1 inhibition can protect against obesity-induced adipose tissue inflammation and improve insulin sensitivity.

[0985] The reduction in inflammation was particularly pronounced in the adipose tissue of mice with FADS1 inhibition. Transcriptomic profiling of EPI and ING WAT in Fads1 KO and Compound A-treated mice revealed downregulation of inflammatory pathways (Figures 4(d), 10(c), 5(a), 13(b)-(c)) and alterations in immune cell populations (Figures 4(b), 8(b)). Decreased PAI-1 and MCP-1 protein levels and monocyte / macrophage markers (Cd14 and Cd68) in the adipose tissue of Fads1 KO and Compound A-treated mice indicated reduced macrophage infiltration and inflammation under FADS1 inhibition (Figures 5(a)-(b), 13(b)-(c), 13(f)). This suggests that FADS1 inhibition can protect against obesity-induced adipose tissue inflammation.

[0986] Obesity leads to a dramatic increase in macrophages in adipose tissue in both humans and mice. Excessive macrophage numbers increase the secretion of inflammatory cytokines, thereby altering insulin signaling and contributing to insulin resistance in obesity. Therefore, reducing macrophage infiltration and subsequent inflammation by inhibiting FADS1 may improve insulin resistance in DIO mice. Insulin sensitivity was improved in Fads1KO and Compound A-treated mice, as demonstrated by improved glucose tolerance and reduced blood glucose and insulin levels (Figures 2(e)-(f), 7, 8(g)), as well as upregulation of insulin signaling genes (Insr, Irs1, Slc2a4) in EPI and ING WAT (Figures 4(d), 14(a), 15(b)-(c)). Reducing inflammation and improving insulin sensitivity by inhibiting FADS1 may help alleviate energy metabolism disorders in the obese state.

[0987] As shown here, the weight loss associated with ablation of FADS1 activity was not attributable to a reduction in food intake, but rather to a shift toward increased lipid oxidation, as indicated by a reduction in RER following genetic and pharmacological FADS1 inhibition (Figures 3(e)-(f), 9(g)-(h)). In addition to the reduction in RER, other hallmarks of increased lipid oxidation were observed, including increased levels of citric acid cycle intermediates in the plasma of Fads1 KO mice (Figure 7). Transcriptome analysis indicated that activation of the PPARα pathway was a mediator of increased lipid oxidation; when FADS1 activity was ablated by genetic deletion or pharmacological inhibition, increased activation of the hepatic PPAR signaling pathway / PPARα / RXRα was observed (Figures 4(c), 10(d)) and increased hepatic expression of fat oxidation genes (Cpt1a, Acox1, Fgf21) (Figures 5(a), 13(a)). In particular, upregulated hepatic expression of the Cyp4a12 gene (Figures 5(a) and 13(a)) strongly suggests PPARα activation, as studies using Ppara KO mice have shown that hepatic expression of the Cyp4a12 gene is dependent on PPARα. Cyp4a expression is extremely sensitive to ligand activation and serves as a marker of PPARα activation. Although the mechanism of PPARα activation is unclear, alterations in PUFA levels and downstream metabolites of FADS1 inhibition may be involved. For example, a high AA / DHA ratio impairs mitochondrial function and reduces PPARα expression, leading to reduced fatty acid oxidation in hepatocytes. The reduced AA / DHA ratio (twofold and fivefold in Fads1 KO mice and Compound A-treated mice, respectively) and FADS1 inhibition may lead to increased fatty acid oxidation (Tables 3.1 and 6.1).

[0988] Table 6.1. PUFA profile in plasma of mice treated with Compound A

[0989]

[0990] In addition to increased fatty acid oxidation, a lipolysis signature (increased expression of Lipe, Mgll, Pnpla2) was observed in EPI WAT of Fads1 KO mice and Compound A-treated mice (Figures 4(d), 5(a), 13(b)). Isolated adipocytes from Fads1 KO mice also had increased isoproterenol-stimulated lipolysis relative to WT adipocytes. Reduced insulin and prostaglandin E2 (PGE2) levels (Figures 2(e), 7, 8(g)), known anti-lipolytic agents, may lead to a significant upregulation of lipolysis genes. Since reduced lipolysis and reduced FA oxidation per kilogram of fat mass are observed in obesity, the concomitant upregulation of adipose lipolysis and hepatic fatty acid oxidation may enhance the mobilization of lipids from WAT to the liver (and possibly other tissues) for oxidation, as a potential mechanism of the anti-obesity effects of FADS1 inhibition. Here, only the liver and two adipose tissue depots are the focus, with EPI WAT representing visceral fat and ING WAT representing subcutaneous adipose tissue. However, since FADS1 is ubiquitously expressed, FADS1 inhibition may affect other metabolically relevant tissues.

[0991] Because AA is a precursor to the endocannabinoids 2-AG and AEA, endocannabinoid concentrations would be expected to be downregulated when AA availability is limited by FADS1 inhibition. This effect was observed in Fads1 KO and Compound A-treated mice (Figures 14(a)-14(b)). Decreased endocannabinoid levels may contribute to the metabolic phenotypes observed in Fads1 KO and Compound A-treated mice, as rimonabant, a selective inverse agonist of cannabinoid receptor 1 (CB1), the receptor for 2-AG and AEA, has been shown to reduce body weight in C57BL / 6DIO mice, with similar phenotypes, including reduced adipose tissue inflammation. However, despite these similarities, differences were also noted. Rimonabant treatment increased physical activity only during the light cycle, whereas FADS1 inhibition or deletion increased physical activity only during the dark cycle (Figures 3(g)-(h), 9(i)-(j)), when mice are more active. Thus, while reduced endocannabinoid levels may partially contribute to the phenotypes observed in Fads1 KO and Compound A-treated mice, the data presented here demonstrate phenotypic differences between rimonabant- and FADS1 inhibitor-treated mice, suggesting that other mechanisms may be involved in driving the improved metabolic phenotype when FADS1 activity is abolished.

[0992] Hepatic PPARα target gene expression and hepatic triglyceride content were monitored in Fads1 KO mice fed a normal diet or HFD (Figures 15(a)-(b)) and in DIO mice treated with Compound A (Figure 15(c)). No significant increase in hepatic triglyceride content was observed. Several groups have reported lower hepatic fat content in Fads1 KO mice. In the livers of FADS1-inhibited mice, a general increase in the expression of hepatic PPARα target genes (e.g., Cpt1a) and downregulation of the lipogenic gene Srebf1 were observed, with Compound A-treated mice showing more pronounced effects compared to Fads1 KO mice (Figures 4(c), 5(a), 10(d), 13(a), 15(e)). No significant increase in hepatic expression of markers of hepatic steatosis was observed in Fads1 KO mice (Figure 15(d)). Plasma liver enzyme alanine aminotransferase (ALT) was reduced in both Fads1 KO and Compound A-treated mice. Furthermore, CP-24879, a dual inhibitor of FADS1 / FADS2 (D5D / D6D), exhibited antisteatotic effects in hepatocytes. Taken together, these data suggest that hepatic steatosis may not be a significant issue with FADS1 inhibition.

[0993] In summary, inhibiting FADS1 activity in DIO mice alleviated obesity and improved the metabolic profile of these mice by reducing inflammation, increasing insulin sensitivity, and lipid oxidation. These improvements were partially due to a decrease in RER and increased physical activity, as measured by indirect calorimetry. Transcriptomic and metabolomic analyses revealed the metabolic status of mice with FADS1 inhibition. The signature of reduced inflammation and increased adiponectin levels observed in epididymal WAT, as well as the upregulation of PPARα-activated fatty acid oxidation genes in the livers of Fads1 KO and Compound A-treated mice, implicate pathways involved in the weight loss and improved metabolic profile mediated by FADS1 inhibition. Furthermore, downregulation of endocannabinoid levels may partially contribute to the anti-obesity mechanism of FADS1 inhibition. Together, these data suggest that FADS1 inhibitors may have potential for treating obesity and related comorbidities.

[0994] FADS1 activity is elevated in obese male mice and humans. Fads1 KO mice are protected from HFD-induced obesity and display improved metabolic parameters. Similarly, chronic administration of the FADS1 inhibitor Compound A to DIO mice significantly reduces body weight and improves the metabolic profile with minimal effects on food consumption. Genetic deletion or inhibition of FADS1 leads to altered lipid profiles, increased lipid oxidation, and reduced inflammation. Multi-omics approaches have revealed insights into the underlying underlying mechanisms, highlighting the differential involvement of metabolic tissues. The mechanism by which FADS1 inhibition combats obesity may involve a simultaneous increase in lipolysis and hepatic lipid oxidation, thereby reducing adiposity.

Claims

1. A method of treating a FADS1-mediated disease or disorder in a subject in need of treatment, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; Wherein, the biological indicator is the ratio of arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA); and If the level of the biological marker in the subject is above the reference level for the biological marker, a therapeutically effective amount of a FADS1 inhibitor compound is administered to the subject.

2. The method of claim 1, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 5:

1.

3. The method of claim 1, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 6:

1.

4. The method of claim 1, wherein the reference level of the biological marker is a ratio of AA to DGLA equal to or at least about 7:

1.

5. A method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological marker is a measured level of linoleic acid, gamma-linoleic acid, adrenic acid, docosapentaenoic acid n-6, alpha-linolenic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosahexaenoic acid, or a combination thereof; and If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject. 6 . The method of claim 5 , wherein when the level of the biological indicator of the subject is higher than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound. 7 . The method of claim 5 , wherein when the level of the biological indicator of the subject is lower than the reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

8. A method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; The biological indicators are the following measurement levels: plasma cholesterol, free cholesterol, total cholesterol, cholesterol ester C20:4, malate, α-ketoglutarate, mannose, glucose, erythro-dihydrosphingosine (d18:0), 5-O-methylsphingosine (d18:1), erythro-sphingosine (d18:1), 3-O-methylsphingosine (d18:1), threo-sphingosine (d18:1), 1-hydroxy-2-amino-(cis, trans)-3,5-octadecadiene, 4-hydroxydihydrosphingosine (t18:0, phytosphingosine), sphingomyelin (d18:1, C23:0), sphingomyelin (d18:1, C24:0), ceramide (d18: 1, C24:0), thromboxane B2, delta-12-prostaglandin D2, prostaglandin E2, prostaglandin D2, 12-hydroxyheptadecatrienoic acid (C17: [5,8,10]3), 14,15-dihydroxyeicosatrienoic acid (C20: cis [5,8,11]3), 11-hydroxyeicosatetraenoic acid (C20: cis [5,8,12,14]4), 13-hydroxyoctadecadienoic acid (13-HODE) (C18: cis [9] trans [11]2), arachidonic acid (C20: cis [5,8,11,14]4), docosahexaenoic acid (C22: cis [4,7,10,13,16,19]6), dihomo- γ-linolenic acid (C20: cis[8,11,14]3), γ-linolenic acid (C18: cis[6,9,12]3), docosapentaenoic acid (C22: cis[7,10,13,16,19]5), eicosapentaenoic acid (C20: cis[5,8,11,14,17]5), docosatetraenoic acid (C22: cis[7,10,13,16]4), stearic acid (C18:0), tryptophan, kynurenic acid, xanthurenic acid, histidine, leucine, isoleucine, valine, 3-hydroxyisobutyric acid, glutamic acid, threonine, cysteine, sarcosine, plasma triglycerides, taurochenodeoxycholic acid, taurocholic acid, lysophosphatidylcholine (C18 :0), lysophosphatidylcholine (C20:4), lysophosphatidylcholine (C17:0), lysophosphatidylethanolamine (C22:5), phosphatidylcholine (C18:0, C22:6), phosphatidylcholine (C18:0, C20:3), phosphatidylcholine (C18:1, C18:2), phosphatidylcholine (C16:1, C18:2), phosphatidylcholine (C18:0, C18:2), phosphatidylcholine (C16:0, C20:5), phosphatidylcholine (C16:0, C16:0), glycerol-3 phosphate, choline plasmalogen (C18, C20:4), inositol, inositol phospholipids, glycerol phosphate, phospholipid fraction, or a combination of the foregoing; as well as If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject. 9 . The method of claim 8 , wherein when the level of the biological indicator of the subject is higher than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

10. The method of claim 8, wherein when the level of the biological indicator of the subject is lower than the reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

11. A method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; The biological indicator is the measured level of differentially expressed genes (DEGs); Among them, the DEGs are Serpinb1a, Gna14, Serpina3m, Hsd11b1, Cyp2c29, Akr1d1, Aldh1a1, Mmp19, Gyp27a1, Tymp, Elovl2, Chkb, H2afj, Tnfaip8l1, Tmem86a, Sel1l3, Agap2, 4833411C07Rik, Elov4, Fat3, Eef1a2, Atp7a, Rgs2, Cenpe, Mfap2, Mlki67, Ctsh, Trub2, Ubc, H6pd, Eepd1, Acss2, Aacs, Gm36827, Man2a2, Nudt18, Plagl1, TM4sf19, Atp6v0d2, Gm20056, Trem2, Il1rn, Mmp12, Cdk18, Efr3b, Tagln2, Lurap1, Cp, I17rb, B230303O12Rik, Cfd, Sult1e1, Tdo2, Cyp2b9, Hao2, Cyp2b13, Cyp2a22, Acnat2, Ildr2, Rpl10a-ps1, Tm6sf2, Fitm1, Lpar1, C6, Cmah, Lbp, Arsg, Glra3, Lad1, A730063M14Rik, Ly6f, Foxi1, Crygc, Defb28, Wfdc9, Phlda2, Aqp6, Gm16411, Adam7, Ppp2r5b, Slc6a7, Gpr50, Ahnak2, S100a6, Mmp14, Htr2b, Hpgds, Gm18537, Pclo, Adrb3, Gm38394, AC154232.2, Cadps, Adgrb2, Gm45470, Sdr9c7, Dsg1c, Slc17a1, Ces1c, Gss, 1810008I18Rik, Tlcd1, Snrk, Akr 1c20, Gm19950, Ttr, Cbfa2t3, Acat2, Pmvk, Abcd3, Acacb, Arhgap27, Rnase9, Wfdc8, Ighv9-2, DerI3 Acap1, Ccl19, Tcf7, Xkrx, Trim46, Zfp369, Zfp871, Pcdhb21, Gm14288, Uprt, Atm, Dchs2, Cped1, Gm 38357 Cck Ckap2 Gm4419 1600015I10Rik Sez6I2 Prnd Gm16702 S100a8 Pcdh12 Malat1 Kcnq1 ot1, ArI4c, Gm42549, Gm37310, Gm37776, Atp2a1, Ckm, Tnnt3, Adipoq, Fabp4, Lep, Retn, Hoxc8, Hox c9, Cebpa, Dgat1, Dgat2, Elovl3, Fas, Scd1, Srebf1, Hilpda, Lipe, Mgll, Plin1, Plin4, Pnpla2, Pnp la3, Ldah, Cs, Gckr, Me1, Pck1, Pdk4, Irs1, Acaa1a, Acads, Acox1, Cpt1a, Cpt1b, Hadhb, Ehhadh, Ppa ra, Ppargc1a, Cyp4a12a, Cyp4a12b, Cyp2e1, Adgre1, Agtr1a, Ccl2, Cd14, Cd68, Il1b, and Tnf, respectively. as well as If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject. 12 . The method of claim 11 , wherein when the level of the biological indicator of the subject is higher than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound. 13 . The method of claim 11 , wherein when the level of the biological indicator of the subject is lower than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

14. The method of claim 11, wherein the DEG is the DEG disclosed in Figures 4a, 5a, 10a, 13a, 13g, 15e, 15f, or a combination of the aforementioned disclosed DEGs.

15. A method of treating a FADS1-mediated disease or disorder in a subject in need thereof, the method comprising: receiving information comparing the level of a biological indicator of a FADS1-mediated disease from the subject to a reference level of the biological indicator; wherein the biological indicator is a measured level of a cell type in the subject, wherein the cell type is adipocytes (Adipo), B cells (Bcell), endothelial cells (Endo), hepatocytes (Hep), Kupffer cells (Kupff), bone marrow cells (Myel), natural killer cells (NK), T cells (Tcell), or a combination thereof; as well as If the subject is determined to be a subject that would benefit from treatment with a FADS1 inhibitor compound by comparing the subject's level of the biological marker to a reference level of the biological marker, a therapeutically effective amount of the FADS1 inhibitor compound is administered to the subject. 16 . The method of claim 15 , wherein when the level of the biological indicator of the subject is higher than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound. 17 . The method of claim 15 , wherein when the level of the biological indicator of the subject is lower than a reference level of the biological indicator, the subject is identified as a subject who will benefit from treatment with the FADS1 inhibitor compound.

18. The method of any one of claims 1 to 17, wherein the FADS1-mediated disease or disorder is obesity, a metabolic disorder, a cardiovascular disorder, diabetes, dyslipidemia, non-alcoholic steatohepatitis (NASH), or a combination of any of the foregoing.

19. The method of any one of claims 1 to 18, wherein the reference level of the biological indicator is the average amount of the biological indicator in a population of healthy subjects.

20. The method of any one of claims 1 to 19, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects not suffering from a FADS1-mediated disease.

21. The method of any one of claims 1 to 20, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects with a body mass index (BMI) greater than or equal to 25.

0.

22. The method of any one of claims 1 to 20, wherein the reference level of the biological marker is the average amount of the biological marker in a population of subjects with a body mass index (BMI) greater than or equal to 30.

0.

23. The method or compound of any one of claims 1 to 22, wherein the level of a biological marker of a FADS1-mediated disease in the subject is quantified using a sample collected from the subject; wherein the sample is blood, plasma, or a tissue biopsy.

24. The method or compound of claim 23, wherein the tissue is adipose tissue or organ tissue.

25. The method or compound of any one of claims 1 to 24, wherein the FADS1 inhibitor compound is: 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-3,7-bis(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-Fluoro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 7-(Trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-chloro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-(methoxymethyl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-cyclopropyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-cyclopropyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2,3-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one; 2,3-Dimethyl-5-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one; 7-ethyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one; 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one; 1,3-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-1H,7H-pyrazolo[1,5-a]pyrimidin-7-one; 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,8H-pyrrolo[1,2-a]pyrimidin-4-one; 2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-(Trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H,6H,7H,8H-pyrrolo[1,2-a]pyrimidin-4-one; 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 8-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-9-(trifluoromethyl)-6,10-diazatricyclo[4.4.0.0 2 , 4 ]Deca-1(10),8-dien-7-one; 6-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-[1,2,4]triazolo[4,3-a]pyrimidin-5-one; 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,3,4]tetrazo[1,5-a]pyrimidin-7-one; 2-methyl-6-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-(Hydroxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-(Hydroxymethyl)-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 2-chloro-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(propan-2-yl)-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione; 1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 3-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5-(trifluoromethyl)-3H,7H-[1,2,4]triazolo[1,5-a]pyrimidin-7-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3]oxazolo[3,2-a]pyrimidin-5-one; 2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 7-Ethoxy-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-(methoxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-methoxy-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 3-chloro-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-(Hydroxymethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-(Hydroxymethyl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,2H,3H,5H-imidazo[1,2-a]pyrimidine-2,5-dione; 2-chloro-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-chloro-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-cyclopropyl-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-chloro-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-dimethyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-Dimethyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-(methoxymethyl)-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-ethyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(2-methoxyethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(propan-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(cyclopropylmethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-(methoxymethyl)-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(2-Hydroxypropyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-dimethyl-6-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(cyclopropylmethyl)-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-[2-(dimethylamino)ethyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(cyclopropylmethyl)-2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-[2-(dimethylamino)ethyl]-2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methoxy-1-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methoxy-1-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[4-(2,2,2-trifluoroethoxy)phenyl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-( 2 H3) methyl-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-( 2 H3) methyl-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(2-hydroxyethyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-5-oxo-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidine-1-carboxylic acid methyl ester; 1-[(2,2-difluorocyclopropyl)methyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-[(3,3-difluorocyclobutyl)methyl]-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(2-hydroxyethyl)-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-[2-(dimethylamino)ethyl]-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(prop-2-yn-1-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-{2-methyl-5-oxo-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-1-yl}acetonitrile; 2-[2-methyl-5-oxo-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-1-yl]acetonitrile; 1-(2-hydroxy-2-methylpropyl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-[2-(1-hydroxycyclopropyl)ethyl]-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-1-[(oxetan-3-yl)methyl]-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-1-(oxetan-3-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1,2-Dimethyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidine-5-thione; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-1-(pyridin-2-yl)-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyrazin-2-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-1-(6-methylpyridin-2-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-1-(1-methyl-1H-pyrazol-4-yl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-3-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-phenyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-(6-chloropyridin-2-yl)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(pyridin-4-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-1-(1H-pyrazol-4-yl)-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 2-(Fluoromethyl)-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 2-[(dimethylamino)methyl]-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-1,2-dimethyl-7-(trifluoromethyl)-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 6-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; 6-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-2-methyl-7-(trifluoromethyl)-5H-[1,3,4]thiadiazolo[3,2-a]pyrimidin-5-one; (2R)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; (2S)-2-methyl-6-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-7-(trifluoromethyl)-2H,3H,5H,6H,7H-[1,3]thiazolo[3,2-a]pyrimidin-5-one; 1-{[(1R)-2,2-difluorocyclopropyl]methyl}-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; 1-{[(1S)-2,2-difluorocyclopropyl]methyl}-2-methyl-7-(trifluoromethyl)-6-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-1H,5H-imidazo[1,2-a]pyrimidin-5-one; (2R)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; (2R)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; (2S)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; (2S)-2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)-1H-pyrazol-1-yl)acetonitrile; (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)acetonitrile; (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenyl)acetonitrile; (4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)acetonitrile; (4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetonitrile; 1-(Chloromethyl)-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-pyrimido[1,2-a][1,3]diazine-2,6-dione; 1-(Fluoromethyl)-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione; 1-(methyl-d3)-7-(4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione; 1-Methyl-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione; 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)-2-methylpropionitrile; 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; 2-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-3-yl)phenoxy)propionitrile; 2-(difluoromethyl)-3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(Difluoromethyl)-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile; 2-(difluoromethyl)-8-methoxy-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(difluoromethyl)-8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(Difluoromethyl)-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(difluoromethyl)-8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(Fluoromethyl)-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile; 2-(Fluoromethyl)-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-(Trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H,6H,7H,9H-pyrimido[2,1-c][1,4]oxazin-4-one; 2,8-dimethoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-cyclopropyl-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-Ethoxy-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-Ethoxy-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 2-ethyl-8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-ethyl-8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 2-ethyl-8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-4,8(1H)-dione; 3-(1-(2,2-difluoropropyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-(3-fluorophenyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-(4-fluorophenyl)-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-{[(1R)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-{[(1S)-2,2-difluorocyclopropyl]methyl}-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-Benzofuran-2-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-cyclopropyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-cyclopropyl-1H-pyrazol-4-yl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-phenyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(1-propyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(2-chloro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(2-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 3-(2-fluoro-4-(trifluoromethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(2-fluoro-6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(3-chloro-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 3-(4-(((1R)-2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(((1S)-2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-((2,2-difluorocyclopropyl)methoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-4,8(1H)-dione; 3-(4-(2,2,2-trifluoroethoxy)phenyl)-2,8-bis(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2-difluoroethoxy)-2-fluorophenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2-difluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2-difluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2,2-difluoropropoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2-fluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2-fluoroethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 3-(4-(2-fluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(2-fluoroethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 3-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)-1H-pyrazol-1-yl)propionitrile; 3-(4-(8-methoxy-4-oxo-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-3-yl)phenyl)propionitrile; 3-(4-(cyclopropylmethoxy)-2-fluorophenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(cyclopropylmethoxy)phenyl)-8-(methyloxy-d3)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(cyclopropylmethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(4-(cyclopropylmethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 3-(4-(difluoromethoxy)phenyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(5-(2,2,2-trifluoroethoxy)-2-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(5-fluoro-6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,9H-pyrimido[2,1-c][1,4]oxazin-4-one; 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-[1-(cyclopropylmethyl)-1H-pyrazol-4-yl]-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 3-[5-iodo-1-(2,2,3,3,3-pentafluoropropyl)-1H-1,2,3-triazol-4-yl]-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-7-fluoro-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-{1-[(2,2-difluorocyclopropyl)methyl]-1H-pyrazol-4-yl}-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methoxy-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 3-{1-[(3,3-difluorocyclobutyl)methyl]-1H-pyrazol-4-yl}-8-methyl-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 3-Fluoro-1-methyl-7-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-8-(trifluoromethyl)-1H,2H,6H-[1,3]diazino[1,2-a]pyrimidine-2,6-dione; 4-Oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile; 4-Oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylic acid; 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carbonitrile; 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; 4-Oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylic acid; 4-Oxo-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-7-carbonitrile; 7-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione; 7-(3-fluoro-4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione; 7-(4-(2,2,2-trifluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione; 7-(4-(2-fluoroethoxy)phenyl)-8-(trifluoromethyl)-2H-pyrimido[1,2-a]pyrimidine-2,6(1H)-dione; 7-(methoxymethyl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7,8-dimethyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one; 7,8-dimethyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 7,9-dimethyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 7-chloro-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 7-chloro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one; 7-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-8-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-chloro-8-methyl-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-cyclopropyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-cyclopropyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-8-hydroxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one; 7-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-8-methoxy-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-Fluoro-8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one; 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 7-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one; 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one; 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 7-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-((1R)-1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((1R)-1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((1S)-1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((1S)-1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((dimethylamino)methyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((methylsulfanyl)methoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((R)-ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((R)-methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((R)-methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((S)-ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((S)-methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-((S)-methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(1,3-oxazol-2-yl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(1-hydroxyethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(1-hydroxyethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(2-hydroxypropan-2-yl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(2-methyl-2-oxetanyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(2-propyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(3-azetidinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(aminomethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(1-azetidinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Chloromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Chloromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-(difluoromethoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(difluoromethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(dimethylamino)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(dimethylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(ethylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Fluoromethoxy)-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Fluoromethoxy)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Fluoromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Fluoromethoxy)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-(Fluoromethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Fluoromethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Hydroxymethyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Hydroxymethyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methoxymethyl)-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylamino)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylamino)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-(methyl-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-(methyloxy-d3)-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Methylsulfanyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(Methylsulfanyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylsulfinyl)-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylsulfinyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-(methylsulfonyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-acetyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-amino-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-amino-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-amino-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-chloro-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-chloro-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-cyclopropyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-cyclopropyl-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-vinyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-Ethoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-ethyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-fluoro-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-Hydroxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(1-(3-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-imidazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(1-(4-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-(4-(3,3,3-trifluoropropyl)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methoxy-2-(trifluoromethyl)-3-[3-(3,3,3-trifluoropropyl)-1,2-oxazol-5-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-[4-(3,3,3-trifluoropropyl)-1H-imidazol-1-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-(trifluoromethyl)-3-[5-(3,3,3-trifluoropropyl)-1,3-thiazol-2-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-2-methyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-(4,4,4-trifluorobutyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-phenyl-1H-pyrazol-3-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-phenyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(1-propyl-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(2-methyl-4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(2-phenyl-1,3-oxazol-5-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(3-phenyl-1,2-oxazol-5-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)-2-(trifluoromethyl)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(2,2,3,3,3-pentafluoropropoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(2,2,3,3-tetrafluoropropoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(trifluoromethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-(trifluoromethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(4-propylphenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(5-propyl-1,2-oxazol-3-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(6-(2,2,2-trifluoroethoxy)-3-pyridinyl)-2-(trifluoromethyl)-4H-pyrimido[1,2-a]pyrimidin-4-one; 8-methoxy-3-(6-propyl-3-pyridyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-1,2,3-triazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-3-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,6-a]pyrimidin-4-one; 8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)-1,3-thiazol-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-[1,3]diazino[1,2-a]pyrimidin-4-one; 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methoxy-3-[2-(2,2,3,3,3-pentafluoropropoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[3-(2,2,3,3,3-pentafluoropropyl)-1,2-oxazol-5-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[4-(2,2,2-trifluoroethoxy)-1,3-thiazol-2-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-[5-(2,2,2-trifluoroethoxy)-1,3-thiazol-2-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-3-{1-[(oxetan-3-yl)methyl]-1H-pyrazol-4-yl}-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methoxy-6-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methyl-2-(trifluoromethyl)-3-[1-(3,3,3-trifluoropropyl)-1H-pyrazol-4-yl]-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methyl-2-(trifluoromethyl)-3-[5-(3,3,3-trifluoropropyl)-1,2,4-oxadiazol-3-yl]-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methyl-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H,6H,7H,8H,9H-pyrimido[1,2-a]pyrazin-4-one; 8-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methyl-3-[2-(2,2,2-trifluoroethoxy)pyrimidin-5-yl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 8-methyl-3-[4-(2,2,2-trifluoroethoxy)phenyl]-2-(trifluoromethyl)-4H-pyrimido[1,2-b]pyridazin-4-one; 9-chloro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 9-Fluoro-8-methoxy-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-4-one; 9-methyl-3-[1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl]-2-(trifluoromethyl)-4H-pyrazino[1,2-a]pyrimidin-4-one; 4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxylic acid methyl ester; Methyl(4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)carbamoyl fluoride; N-(4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetamide; N-(4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidin-8-yl)acetamide; N,N-dimethyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; N-ethyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; N-methyl-4-oxo-3-(1-(2,2,3,3,3-pentafluoropropyl)-1H-pyrazol-4-yl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; or N-methyl-4-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)-2-(trifluoromethyl)-4H-pyrido[1,2-a]pyrimidine-8-carboxamide; or A pharmaceutically acceptable salt of any of the foregoing.

26. The method or compound of any one of claims 1 to 25, wherein the FADS1 inhibitor compound is the free base.

27. A method for measuring the ratio of arachidonic acid (AA) to dihomo-gamma-linolenic acid (DGLA) in a subject; wherein the AA is isotopically labeled and the DGLA is isotopically labeled; wherein the AA to DGLA ratio is measured by administering a dose of labeled DGLA to the subject and then measuring the ratio of labeled AA to labeled DGLA.

28. The method of claim 27, wherein the labeled DGLA comprises an enriched isotope 13 DGLA of C.

29. The method of claim 28, wherein the labeled DGLA comprises isotopes having equal to or at least five carbon position enrichments. 13 DGLA of C atoms.

30. The method of any one of claims 27 to 29, wherein the labeled DGLA comprises a label at one or more or all of the carbons indicated below with "*":

31. The method of any one of claims 27 to 30, wherein a dose of a FADS1 inhibitor compound is administered to the subject before, simultaneously with, or after administration of a labeled dose of DGLA to the subject.

32. A compound represented by the following structure: in Each "*" symbol indicates an isotope that can be enriched 13 The location of C; and At least one "*" position is isotope enriched 13 C.

33. The compound of claim 32, wherein at least five "*" positions are isotopically enriched 13 C.

34. A compound as described in any one of claims 32 to 33, wherein the enrichment factor of each isotopically enriched position is equal to or at least 1000.

35. A method for producing a compound represented by the following structure: The method comprises making the following compound Reacts with KC*N; in Each "*" symbol indicates an isotope that can be enriched 13 C position and at least one "*" position is isotopically enriched 13 C; and wherein X is a halogen.

Images