Therapeutic formulations for kidney disease comprising repagermanium or propagermanium and / or an AT1R blocker

Abstract

The present invention relates to therapeutic formulations adapted to deliver a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium, therapeutic formulations adapted to deliver a therapeutically effective daily dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone, and combinations thereof. These formulations and combinations are used for the treatment of renal disease.

Description

Technical Field

[0001] Technical Field The present disclosure relates to therapeutic formulations containing repagermanium or propagermanium and methods of using the same, as well as therapeutic formulations containing an AT1R blocker and methods of using the same. The repagermanium or propagermanium can be delivered in a therapeutic combination regimen with an AT1R blocker.

Background Art

[0002] Background Art Therapeutic methods (where necessary, in a therapeutic combination regimen) for diseases such as kidney diseases and respiratory diseases based on chemokine receptor 2 (CCR2) pathway inhibitors or AT1R blockers are known.

[0003] Repagermanium or propagermanium can be used as a CCR2 pathway inhibitor. Repagermanium and propagermanium have previously been described in the literature as polymer compounds. The prodrug, 3-[(2-carboxyethyl-oxogermyl)oxy-oxogermyl]propanoic acid, exhibits polymorphism and can exist in various solid forms. However, the inventors have not observed any pharmacological effects of repagermanium polymerization during the production or testing of the prodrug.

[0004] Examples of the polymorphism of 3-[(2-carboxyethyl-oxogermyl)oxy-oxogermyl]propanoic acid include repagermanium, propagermanium, proxigermanium, Ge-132, germanium sesquioxide, 2-carboxyethylgermasesquioxane, SK-818, and bis(2-carboxyethylgermanium) sesquioxide.

[0005] The two most highly characterized solid forms, repagermanium and propagermanium, share a common core structure (Mizuno, et al, 2015). These two active pharmaceutical ingredients have different international common names (INNs), with repagermanium having INN #6259 and propagermanium having INN #6260. They consist of different packings of monomer molecules, 3-[(2-carboxyethyl-oxogermyl)oxy-oxogermyl]propanoic acid, varying in the structural units coordinated by H bonds. X-ray diffraction revealed that repagermanium has an infinite sheet structure, clearly distinguishable from the propagermanium polymer ladder structure of chained 8-membered rings composed of Ge-O bonds. In solid form, repagermanium and propagermanium are different; however, in solution, they are identical due to the dissolution of their structural units.

[0006] Angiotensin type I receptors (AT1R, AT1R, angiotensin II receptor type 1) are G protein-coupled receptors. Regarding their effects on AT1R blockade, AT1R blockers (also known as ARBs) have traditionally been considered therapeutically interchangeable, regardless of structural differences.

[0007] However, dosing regimens for repagermanium or propagermanium and AT1R blockers have not yet been clearly established for various diseases. There is a need to establish appropriate dosing regimens for repagermanium or propagermanium and AT1R blockers; or at least to provide new dosing regimens that complement previously known dosing regimens. This disclosure seeks to provide improved or alternative dosing regimens for repagermanium or propagermanium and AT1R blockers. The above-mentioned repagermanium or propagermanium may be delivered in combination regimens with AT1R blockers.

[0008] The prior consideration of the background technology is intended solely to facilitate understanding of this disclosure. That consideration does not constitute an acknowledgment or assertion that any of the referenced materials were / were part of common knowledge at the time of the priority date of this application. [Overview of the project] [Means for solving the problem]

[0009] Summary of the Invention This disclosure provides a therapeutic formulation adapted to deliver a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

[0010] The disclosure further provides a therapeutic formulation adapted to deliver a therapeutically effective daily dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone for the AT1R blocker.

[0011] The disclosure further provides a therapeutic formulation adapted to deliver a therapeutically effective daily dose of an AT1R blocker having (a) a therapeutically effective amount of repagermanium or propagermanium; and (b) an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0012] If necessary, the above daily dose of repagermanium or propagermanium is approximately 200 mg to 280 mg.

[0013] If necessary, the above AT1R blocker is selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan.

[0014] If necessary, the above daily dose of AT1R blockers is approximately 2 mg to 800 mg.

[0015] This disclosure further provides a dosage regimen that includes a step of administering a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

[0016] The disclosure further provides a dosing regimen comprising the step of administering a therapeutically effective daily dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0017] The disclosure further provides a dosing regimen comprising the steps of (a) administering a therapeutically effective amount of repagermanium or propagermanium; and (b) administering a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0018] The Disclosure further provides a method for treating, improving or preventing kidney disease, the method comprising the step of administering to a subject a therapeutic formulation adapted to deliver (a) about 200 mg to 280 mg of repagermanium or propagermanium per day.

[0019] The disclosure further provides a method for treating, improving or preventing kidney disease, the method comprising the step of administering to a subject a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0020] The Disclosure further provides a method for treating, improving or preventing kidney disease, the method comprising the steps of administering to a subject (a) a therapeutically effective amount of repagermanium or propagermanium per day; and (b) a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0021] The present disclosure further provides for the use of a daily dose of about 200 mg to 280 mg of repagermanium or progermanium for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0022] The present disclosure further provides for the use of a daily dose of about 200 mg to 280 mg of repagermanium or progermanium for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0023] The present disclosure further provides for the use of a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is the same as or greater than the affinity of the AT1R blocker for only AT1R, for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0024] The present disclosure further provides for the use of a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is the same as or greater than the affinity of the AT1R blocker for only AT1R, for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0025] The present disclosure further provides for the use of (a) a therapeutically effective amount of repagermanium or progermanium, and (b) a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is the same as or greater than the affinity of the AT1R blocker for only AT1R, for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0026] The present disclosure further provides for the use of (a) a therapeutically effective amount of repagermanium or progermanium, and (b) a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is the same as or greater than the affinity of the AT1R blocker for only AT1R, for the manufacture of a formulation for treating, ameliorating or preventing kidney disease in a subject.

[0027] The disclosure further provides therapeutically effective amounts of repagermanium or propagermanium for use in formulations for the treatment, improvement or prevention of renal disease in subjects, wherein the repagermanium is administered to the subject simultaneously or sequentially with a therapeutically effective amount of AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of an AT1R blocker for AT1R alone.

[0028] The disclosure further provides therapeutically effective amounts of AT1R blockers having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone of an AT1R blocker, for use in formulations for the treatment, improvement or prevention of renal disease in subjects, wherein at least one AT1R blocker is administered to the subject simultaneously or sequentially with repagermanium or propagermanium.

[0029] The Disclosure further provides a kit for the treatment or prevention of kidney disease in a subject, the kit comprising (a) a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium; and (b) instructions for use.

[0030] The disclosure further relates to a kit for the treatment or prevention of kidney disease in a subject, the kit comprising (a) a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone; (b) Provide a kit that includes instructions for use.

[0031] The Disclosure further provides a kit for the treatment or prevention of kidney disease in a subject, the kit comprising: (a) a therapeutically effective amount of repagermanium or propagermanium; (b) a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone; and (c) instructions for use.

[0032] If necessary, the above-mentioned repagermanium or propagermanium will be supplied in a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

[0033] If necessary, the above AT1R blocker is selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan.

[0034] The subjects to be treated, if necessary, are human mammals.

[0035] If necessary, the above-mentioned kidney diseases are those associated with proteinuria.

[0036] If necessary, the above kidney diseases are selected from a list that includes focal segmental glomerulosclerosis (FSGS; including idiopathic (primary) FSGS, secondary FSGS, hereditary FSGS, and any other cause of FSGS), chronic kidney diseases including chronic kidney disease caused by fibrous disorders of the kidney, immunoglobulin A nephropathy (IgAN), Alport syndrome, and diabetic nephropathy, renal insufficiency (diabetic and non-diabetic), and renal failure conditions including diabetic nephropathy, glomerulonephritis, scleroderma, glomerulosclerosis, proteinuria of primary kidney disease, and renovascular hypertension.

[0037] If necessary, the AT1R blocker mentioned above is not EXP3174. Brief explanation of the drawing Further features of this disclosure are better described in the following description of some non-limiting embodiments thereof. This description is included solely for illustrative purposes of this disclosure and should not be understood as a limitation to its broad gist, disclosure, or description of the disclosure as set forth above. This description is made with reference to the accompanying drawings. [Brief explanation of the drawing]

[0038] [Figure 1] Figure 1 is a graph showing the effect of various doses of repagermanium on the effectiveness of treatment. The effectiveness of repagermanium doses shows a "bell-shaped" inhibition curve. [Figure 2] Figure 2 is a correlation plot graph comparing the efficacy of ARBs using AT1R alone to the efficacy of inhibiting G protein dissociation from the CCR2-AT1R complex when co-stimulated. Cand; candesartan, CandC; candesartan cilexetil, Epro; eprosartan, EXP; EXP3174, Irb; irbesartan, Fim; fimasartan*, Los; losartan, Olm; olmesartan, OlmM; olmesartan medoxomil, Spar; sparnentan, Tel; telmisartan, Val; valsartan. [Figure 3] Figure 3 is a graph of the statistical analysis of the EXP3174 efficacy (pIC50) of AT1R alone compared to the efficacy in inhibiting G protein dissociation from the co-stimulated CCR2-AT1R complex. **, p<0.01 (measured by a one-sample t-test). [Figure 4] Figure 4 is a graph showing the effects of ARB, CCR2, or ARB+CCR2 inhibition on the dissociation of G protein (Gαi1-luciferase) from the CCR2-AT1R complex under simultaneous stimulation. [Figure 5]Figure 5 is a correlation plot graph comparing the efficacy of ARBs with AT1R alone to the efficacy of inhibiting G protein dissociation from the CCR2-AT1R complex co-stimulated in the presence of repagermanium*. Cand; candesartan, CandC; candesartan cilexetil, Epro; eprosartan, EXP; EXP3174, Fim; fimasartan*, Irb; irbesartan, Los; losartan, Olm; olmesartan, OlmM; olmesartan medoxomil, Spar; sparnentan, Tel; telmisartan, Val; valsartan. [Modes for carrying out the invention]

[0039] Description of the Invention Detailed description of the invention While CCR2 inhibitors and / or AT1R blockers have been used in a variety of therapeutic contexts, dosing regimens that provide effective therapeutic outcomes remain largely unknown.

[0040] formulation This disclosure provides a therapeutic formulation comprising (i) repagermanium or propagermanium adapted to be delivered at approximately 200 mg to 280 mg per day; (ii) an AT1R blocker having the same or greater affinity for the CCR2-AT1R complex as the affinity for AT1R alone; or (iii) repagermanium or propagermanium adapted to be delivered at approximately 200 mg to 280 mg per day and an AT1R blocker having the same or greater affinity for the CCR2-AT1R complex as the affinity for AT1R alone.

[0041] Repagermanium or propagermanium preparations The inventors have surprisingly found that the dose of repagermanium or propagermanium required for therapeutic efficacy falls within a narrow range of dosages and treatment regimens.

[0042] Repagermanium and propagermanium block pathways associated with MCP-1-induced migration, monocyte activation, and chemotactic migration. Repagermanium and propagermanium further block the function of chemotactic cellular structures such as microfilaments, microtubules, and intermediate filaments, and surface proteins (e.g., glycosylphosphatidylinositol (GPI) anchored proteins, and more specifically, CD55, CD59, and CD16).

[0043] The INN of repagermanium is INN #6259, while that of propagermanium is INN #6260. Propagermanium is used as an active agent for chronic hepatitis and has been shown to specifically inhibit MCP-1-mediated chemotactic migration of monocytes in vitro through a mechanism that appears to require glycosylphosphatidylinositol (GPI) anchored proteins (e.g., CD55, CD59, and CD16) (Yokochi, S. (2001) Journal of Interferon and Cytokine Research 21:389-398). Further names for propagermanium and / or repagermanium include 3-[(2-carboxyethyl-oxogermyl)oxy-oxogermyl]propanoic acid, proxigermanium, Ge-132, bis(2-carboxyethylgermanium)sesquioxide (CEGS), 2-carboxyethylgermasesquioxane, SK-818, organogermanium, germanium sesquioxide, 3,3'-(1,3-dioxo-1,3-digermanoxanediyl)bispropanoic acid, 3-oxygermylpropionic acid polymer, and poly-trans-(2-carboxyethyl)germasesquioxane. In this disclosure, the term “repagermanium” is defined to include “propagermanium.”

[0044] The inventors found that, for example, a dose of 80 mg twice daily resulted in an excessive decrease in the blood concentration of repagermanium or propagermanium before the next dose, thus failing to provide a therapeutic effect. Furthermore, contrary to expectations, a dose of, for example, 140 mg twice daily resulted in a reduction in therapeutic effect. Therefore, the therapeutic effect of repagermanium or propagermanium is maximized at moderate doses. This general trend can be seen in Figure 1.

[0045] This disclosure therefore provides therapeutic formulations adapted to deliver a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

[0046] The disclosure further provides a dosing plan that includes a step of administering a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

[0047] The above-mentioned repagermanium or propagermanium can be adapted to be delivered at approximately 220 mg to 260 mg / day, or approximately 240 mg / day if necessary.

[0048] Preparations containing repagermanium or propagermanium may be delivered twice daily (BID). Therefore, these preparations may be used in dosing regimens that include administering doses of approximately 100 mg to 140 mg of repagermanium or propagermanium twice daily. Dosing regimens of three or more doses per day are considered, but these are unlikely to be tolerable or adherent to by patients receiving the treatment. If the above dosing is performed three or more times per day, the total daily dose should still be approximately 200 mg to 280 mg / day, and if necessary, approximately 240 mg / day.

[0049] When the above-mentioned repagermanium or propagermanium is being administered to children under 12 years of age, an adjusted dose may be given to achieve the same exposure and pharmacokinetic outcomes as the adult dose. The adjusted dose may be determined based on body weight. The adult dose of approximately 200-280 mg of repagermanium / day is based on an average adult weighing 70 kg. For children under 12 years of age, the dose may be calculated based on their body weight relative to 70 kg.

[0050] The therapeutic formulations comprising repagermanium or propagermanium of this disclosure include pharmacokinetic data, in particular the half-lives of repagermanium and propagermanium in the body and the C of repagermanium and propagermanium. max It is being developed based on [the following].

[0051] This specific dose and exposure of repagermanium or propagermanium is required for peak therapeutic efficacy based on reduced chemotaxis of CCR2-expressing cells.

[0052] Angiotensin type 1 receptor blocker activity Surprisingly, the inventors found that while AT1R blockers were previously considered interchangeable, there are some exceptions based on the interaction between AT1R blockers and the CCR2-AT1R complex.

[0053] The angiotensin type I receptor (AT1R, AT1R, angiotensin II receptor type 1) is a G protein-coupled receptor.

[0054] The term "angiotensin type 1 receptor inhibitor" (also known as angiotensin receptor blockers or ARBs) is understood to mean drugs or compounds that can inhibit or partially inhibit the activation of AT1R. This includes antagonists, inverse agonists, and negative allosteric modifiers of AT1R.

[0055] The term "AT1R blocker" includes pharmaceutically acceptable salts of AT1R blockers. Alternatively, the above-mentioned AT1R blocker may be an antibody blocker of AT1R.

[0056] For example, the above AT1R blockers may be selected from the group including: irbesartan (e.g., Avapro®), eprosartan (e.g., Tebeten®), losartan (e.g., Cozar®), valsartan (e.g., Diovan®), telmisartan (e.g., Micardis®), candesartan (e.g., Atacand®), olmesartan (e.g., Benicar®), azilsartan (e.g., Idarbi®), spalsentan (e.g., Filspari), fimasartan (e.g., Kanarb®), and ZD-7115. For example, the angiotensin receptor inhibitor may be irbesartan or valsartan.

[0057] Taylor et al. (2011) (Journal of the American Society of Hypertension 13(9):677-686) reviewed angiotensin II AT1 receptor blockers (also known as AT1R blockers or ARBs) and noted that "there are no significant differences in efficacy or other clinical properties among older drugs in this class." It is noted that differences in properties relate to interactions with other pathways but do not appear to be related to core angiotensin II AT1 receptor blockade. Taylor reports that "there are some pharmacological differences among ARBs, and some drugs in this class have special effects unrelated to angiotensin II receptor blockade, but there is currently no strong evidence that these translate to specific protection against target organ damage from hypertension." Therefore, with respect to their effects on AT1R blockade, ARBs are functionally interchangeable despite structural differences.

[0058] As noted in Taylor's first paragraph, ARBs first became commercially available in the 1990s, and eight specific ARBs are cited as being available in the United States. Despite the long-term use of this class of drugs, physicians generally seem to treat them as a group that is interchangeable with one another. There are certain specific nuances in the precise pharmacology of some of the above groups, but these do not appear to be related to their angiotensin II AT1 receptor activity. Furthermore, all pharmacological products behave subtly differently in some patients, but despite some variability in the subgroups, they remain generally useful in treatment.

[0059] The AT1R blockers described above inhibit or partially inhibit AT1R as needed. The AT1R blockers do not contain therapeutic agents such as angiotensin-converting enzyme inhibitors (ACEi) that directly interact with AT1R, acting upstream of AT1R to prevent the production of functional ligands, but do not interact with AT1R itself.

[0060] The term "AT1R blocker" includes pharmaceutically acceptable salts of AT1R blockers. Alternatively, the above-mentioned AT1R blocker may be an antibody blocker of AT1R.

[0061] For example, the above angiotensin receptor inhibitor may be irbesartan. Irbesartan is an angiotensin type I receptor antagonist also known as 2-butyl-3-({4-[2-(2H-1,2,3,4-tetrazole-5-yl)phenyl]phenyl}methyl)-1,3-diazaspiro[4.4]nona-1-en-4-one. The above angiotensin receptor inhibitor may be spalsentan. Sparsentan is also known as 2-[4-[(2-butyl-4-oxo-1,3-diazaspiro[4.4]nona-1-en-3-yl)methyl]-2-(ethoxymethyl)phenyl]-N-(4,5-dimethyl-1,2-oxazole-3-yl)benzenesulfonamide and is a combined selective antagonist of the angiotensin II type 1 receptor (AT1R) and endothelin A receptor.

[0062] The CCR2-AT1R complex has been previously identified as a target for the treatment of kidney and other diseases. We have previously demonstrated constitutively high inhibition of inositol phosphate signaling by the AT1R-CCR2 heteromer with ARBs. They have also shown that the CCR2-AT1R complex results in the activation of Gαi protein and the recruitment of β-arrestin.

[0063] However, it remains unknown whether ARBs maintain their efficacy in inhibiting Gαi protein activation or mobilizing β-arrestin when acting on the CCR2-AT1R complex compared to AT1R alone. This is important because ARB-mediated AT1R inhibition is required for clinical efficacy. In Gαi protein dissociation assays, the efficacy of ARBs on AT1R alone generally showed a good correlation with the efficacy of ARBs on the CCR2-AT1R complex when co-stimulated.

[0064] Surprisingly, however, compound EXP3174 was found to exhibit the predicted efficacy at the lower CCR2-AT1R complex in the presence of AT1R alone. Therefore, the AT1R blocker EXP3174 does not function as effectively as other AT1R blockers in the presence of the AT1R-CCR2 complex. This indicates that the affinity between ARBs and the CCR2-AT1R complex is not clear. ARBs that exhibit lower efficacy at the CCR2-AT1R complex compared to AT1R alone may have reduced clinical efficacy.

[0065] The AT1R blocker EXP3174, mentioned above, is a metabolite of losartan, which is widely used in research, and has been reported to be significantly more potent than losartan in blocking the angiotensin II-induced cellular response (Sachinidis et al (1993) EXP3174, a metabolite of losartan (MK 954, DuP 753) is more potent than losartan in blocking the angiotensin II-induced responses in vascular smooth muscle cells J Hypertens 11(2):155-62). EXP3174 was used as an experimental AT1R blocker in combination with a CCR2 pathway inhibitor in WO2012094703.

[0066] This disclosure therefore provides a formulation comprising a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0067] For the purposes of this disclosure, the same or higher affinity of an AT1R blocker to the CCR2-AT1R complex described above is considered to be AT1R blocker affinity (AT1R locker affinity) when there is a loss of 3.5 times or less in affinity at the CCR2-AT1R complex compared to affinity at AT1R.

[0068] The disclosure further provides a dosing regimen comprising the step of administering a therapeutically effective daily dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone.

[0069] If necessary, the AT1R blocker is selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan. In one instance, the AT1R blocker is not EXP3174.

[0070] If the AT1R blocker has a lower affinity for the CCR2-AT1R complex than AT1R alone, the experimental results regarding administration establishment may not be transferable to a clinical setting. The therapeutic dose of an AT1R blocker with a lower affinity for the CCR2-AT1R complex than AT1R alone is likely to be higher than that predicted and established in experimental settings using only AT1R alone. This is because, in the subject's body, AT1R is delivered in the CCR2-AT1R complex. Therefore, a larger amount of the AT1R blocker with a lower affinity for the CCR2-AT1R complex than AT1R alone is required in the body to achieve the same therapeutic effect.

[0071] If necessary, the above AT1R blockers are supplied in doses of approximately 2 mg to 800 mg / day, and are supplied in doses of 1 or more. For example, the above AT1R blockers may be supplied in doses of approximately 5 mg to 320 mg / day. For example, the above AT1R blocker is irbesartan, administered in doses of approximately 75 mg, 150 mg, or 300 mg per day, and is supplied in doses of 1 or more. Alternatively, the above AT1R blocker is valsartan, administered in doses of approximately 80 mg, 160 mg, or 320 mg per day, and is supplied in doses of 1 or more.

[0072] As discussed above, AT1R blockers are functionally interchangeable, as described by Taylor et al. (2011), but the dosages differ for various active agents. Those skilled in the art will use their knowledge of dosage ranges for each AT1R blocker to establish a dosage suitable for use in this disclosure. Table 1: Recommended daily dose of AT1R blockers [Table 1-1] [Table 1-2]

[0073] Combination formulations While not bound by any theory, it is suggested that inhibition of the CCR2 pathway and AT1R signaling is preferable for clinical efficacy. For this reason, a combination of a CCR2 pathway inhibitor and an ARB having the same or greater affinity for the CCR2-AT1R complex as AT1R alone is preferred.

[0074] Examples of such combinations include repagermanium or propagermanium in combination with the ARBs candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan, as well as the dual ARB endothelin receptor antagonist spalsentan. The above combinations do not include repagermanium or propagermanium in combination with EXP3174.

[0075] This disclosure will be furthered, a) A therapeutically effective amount of repagermanium or propagermanium; and b) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. To provide a formulation adapted for delivery.

[0076] This disclosure will be furthered, a) A daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium; and b) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. To provide a formulation adapted for delivery.

[0077] This disclosure will be furthered, a) Approximately 200 mg to 280 mg of repagermanium or propagermanium; b) A therapeutically effective dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. The present invention provides a dosing regimen that includes the step of administering a daily dose of [the substance].

[0078] This disclosure will be furthered, c) Approximately 200 mg to 280 mg of repagermanium or propagermanium; d) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. The present invention provides a dosing regimen that includes the step of administering a daily dose of [the substance].

[0079] The AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone may be selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan. In one aspect, the AT1R blocker is not EXP3174.

[0080] For the purposes of this disclosure, the same or greater affinity of an AT1R blocker to the CCR2-AT1R complex is considered to be AT1R blocker affinity when there is a loss of 3.5 times or less in affinity to the CCR2-AT1R complex compared to affinity to AT1R.

[0081] The therapeutically effective dose of the above-mentioned AT1R blockers may be approximately 2 mg to 800 mg per day.

[0082] The above-mentioned pharmaceutical preparations may, as necessary, contain additives, solvents, carriers, and other pharmaceutically acceptable components.

[0083] The term “component” means either the AT1R blocker or repagermanium as used herein in the context of the pharmaceutical formulations of this disclosure.

[0084] As used herein, the term “inhibit” means a reduction that is below the detection limit compared to a reference. The phrase includes blocking, delaying, or interfering with an action in order to prevent an undesirable outcome.

[0085] As used herein, the term “partially inhibit” means any reduction within the limits of detection when compared to a reference. The foregoing includes blocking, delaying, or interfering with an action in order to prevent an undesirable outcome.

[0086] Both the repagermanium and the AT1R blocker described above may be the same active agent, such as a bispecific antibody.

[0087] Both the AT1R inhibitors and repagermaniums described above may be pharmaceutically acceptable salts of their respective active agents. Pharmaceutically and veterinarily acceptable salts include those that maintain the biological efficacy and properties of the compounds disclosed herein and are not biologically or otherwise undesirable. In many cases, the compounds disclosed herein may form acid salts and / or base salts due to the presence of amino groups and / or carboxyl groups or similar groups. Acceptable base addition salts may be prepared from inorganic and organic bases. Examples of salts obtained from inorganic bases include, but are not limited to, salts of sodium, potassium, lithium, ammonium, calcium, and magnesium. Salts obtained from organic bases include primary, secondary, and tertiary amines (for example, merely illustrative examples: alkylamines, dialkylamines, trialkylamines, substituted alkylamines, di(substituted alkyl)amines, tri(substituted alkyl)amines, alkenylamines, dialkenylamines, trialkenylamines, substituted alkenylamines, di(substituted alkenyl)amines, tri(substituted alkenyl)amines, cycloalkylamines, di(cycloalkyl)amines, tri(cycloalkyl)amines, substituted cycloalkylamines, disubstituted cycloalkylamines, trisubstituted cycloalkylamines, cycloalkenylamines, di(cycloalkenyl)amines, tri(cycloalkenyl)amines, Examples include, but are not limited to, salts of substituted cycloalkenylamines, disubstituted cycloalkenylamines, trisubstituted cycloalkenylamines, arylamines, diarylamines, triarylamines, heteroarylamines, diheteroarylamines, triheteroarylamines, heterocyclic amines, diheterocyclic amines, triheterocyclic amines, mixed diamines, and triamines, wherein at least two of the substituents on the amine are different and selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic, etc. Amines in which two or three substituents combine with an amino nitrogen to form a heterocyclic or heteroaryl group are also included.

[0088] Pharmaceutically and veterinarily acceptable acid addition salts can be prepared from inorganic and organic acids. Examples of inorganic acids that can be used include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of organic acids that can be used include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid.

[0089] Pharmaceutically or veterinarily acceptable salts of compounds useful in this disclosure can be synthesized by conventional chemical methods from parent compounds containing basic or acidic moieties. Generally, such salts can be prepared by reacting the free acidic or basic form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile may be used. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th edition, Mack Publishing Company, Easton, Pa. (1985), p. 1418 (the disclosure of which is incorporated herein by reference).Examples of such acceptable salts include iodide salts, acetates, phenylacetates, trifluoroacetates, acrylates, ascorbic acid salts, benzoates, chlorobenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, methylbenzoates, o-acetoxybenzoates, naphthalene-2-benzoates, bromides, isobutyrates, phenylbutyrates, γ-hydroxybutyrates, β-hydroxybutyrates, butyn-1,4-dioate, hexyn-1,4-dioate, hexyn-1,6-dioate, caproates, caprylates, chlorides, cinnamates, citrates, decanoates, formates, fumarates, glycolates, heptanoates, hippurates, lactates, malates, maleates, hydroxymaleates, malons, mandelates, and mesylates. These include salts, nicotinates, isonicotinates, nitrates, oxalates, phthalates, terephthalates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, propiolates, propionates, phenylpropionates, salicylates, sebacinates, succinates, suberates, sulfates, bisulfates, pyrosulfates, sulfites, bisulfites, sulfonates, benzenesulfonates, p-bromophenylsulfonates, chlorobenzenesulfonates, propanesulfonates, ethanesulfonates, 2-hydroxyethanesulfonates, methanesulfonates (merhanesulfonates), naphthalene-1-sulfonates, naphthalene-2-sulfonates, p-toluenesulfonates, xylenesulfonates, tartrates, etc.

[0090] Condition or disease In general, a range of diseases associated with chemokines can be treated by the methods of this disclosure. This includes diseases associated with increased or decreased production of chemokines and / or increased or decreased cellular responsiveness to chemokines. It should also be understood that chemokine-related diseases also mean diseases or conditions in which chemokine receptors exhibit abnormal characteristics and are targeted by specific pathogens or pharmacological interventions.

[0091] Diseases associated with chemokines may be inflammatory conditions, or conditions resulting from or related to inflammation. These inflammatory conditions, or conditions resulting from or related to inflammation, may include kidney diseases.

[0092] The above-mentioned inflammatory conditions, or conditions resulting from or associated with inflammation that should be treated, improved or prevented, may be kidney diseases. For example, the above-mentioned kidney diseases are those associated with proteinuria. The above-mentioned kidney diseases may be selected from a list that includes focal segmental glomerulosclerosis (FSGS; including idiopathic (primary) FSGS, secondary FSGS, hereditary FSGS, and any other cause of FSGS), chronic kidney diseases including chronic kidney diseases caused by fibrous disorders of the kidneys, immunoglobulin A nephropathy (IgAN), Alport syndrome, and diabetic nephropathy, renal insufficiency (diabetic and non-diabetic), and renal failure conditions including diabetic nephropathy, glomerulonephritis, scleroderma, glomerulosclerosis, proteinuria of primary kidney disease, and renovascular hypertension.

[0093] The term "prevention" includes administering the formulation of this disclosure to a subject who has symptoms indicating that the subject is at risk of developing the kidney disease described above (but has not been diagnosed with the kidney disease at the time of treatment). For example, the subject may have elevated blood creatine or proteinuria, but has not yet been diagnosed with kidney disease.

[0094] The term "improvement" includes the administration of the formulations of this disclosure to subjects who have a history of kidney disease and are at risk of developing the previous disease condition.

[0095] Measurement of inhibition (i) the inhibition or partial inhibition of the CCR2 pathway and / or AT1R caused by the above-mentioned repagermanium or propagermanium, (ii) the above-mentioned AT1R blocker, or (iii) a combination of both repagermanium or propagermanium and the above-mentioned AT1R blocker may be measured using the in vitro methods described herein, including but not limited to biochemical or cytoassays for: Evaluation of in vitro chemotactic migration of CCR2-expressing neutrophils and other cells (e.g., as known in the art), as well as inositol phosphate production, extracellular regulatory kinase (ERK) phosphorylation, cAMP production, unlabeled techniques (e.g., using impedance, photorefraction or charge redistribution), G protein coupling using proximal reporter systems or other approaches, β-arrestin recruitment or mediated signaling, measurement of transcription factor-based reporter systems, visualization by microscopy using fluorescent labeling, use of antibodies to assess receptor cell localization (e.g., enzyme-linked immunosolvent assays), use of bioluminescence resonance energy transfer to assess cell localization and trafficking, and fluorescence-activated cell sorting.

[0096] Inhibition or partial inhibition of the CCR2 pathway and / or AT1R caused by (i) the repagermanium or propagermanium described above, (ii) the AT1R blocker described above, or (iii) a combination of both the repagermanium or propagermanium described above and the AT1R blocker described above, may be measured using in vivo methods including, but are not limited to,: measurement of cellular and cytokine content of pulmonary exudate, pulmonary function measurements including physical volume of pulmonary function using spirometry-based tests, or pulmonary function output measured using blood gas measurements or other biochemical measures, or improvement in functional benefit including clinical benefit measured by quantitative methods such as gait testing or qualitative methods such as patient-reported outcome assessment. Inhibition or partial inhibition may be indicated by qualitative improvement in pulmonary structure as measured by one or more of the endpoints described above.

[0097] In one embodiment, the overall efficacy of the pharmaceutical formulation is greater than the efficacy of either the AT1R blocker or the repagermanium or propagermanium component when administered without any administration of the other component. Therefore, the combined formulation can be administered in a single dose (including a subtherapeutic dose) or at a lower frequency than when either of the two components can be administered as a single compound.

[0098] Where necessary, the overall efficacy of the above pharmaceutical formulation is greater when compared to the sum of the efficacy of any one component of the AT1R blocker and repagermanium or propagermanium when administered without any other component. Synergistic effects in efficacy may be observed when the AT1R blocker and repagermanium or propagermanium are administered simultaneously or sequentially.

[0099] Alternatively, the overall efficacy of the above pharmaceutical formulation may be equal to the sum of the efficacy of any one component of the AT1R blocker and the repagermanium or propagermanium when administered without any administration of the other components. In a further alternative embodiment, an additive effect in efficacy is observed when the AT1R blocker and the repagermanium or propagermanium are administered simultaneously or sequentially.

[0100] In further alternatives, the overall efficacy of the above pharmaceutical formulation is less than the sum of the efficacy of any one component of the AT1R blocker and the repagermanium or propagermanium when administered without any other component. In further embodiments, the combined efficacy is less than the sum of the efficacy of each component of the AT1R blocker and the repagermanium or propagermanium when administered without any other component, but the treatment provides greater efficacy compared to a single treatment of the AT1R blocker or the repagermanium or propagermanium administered alone.

[0101] If necessary, the two components described above may be administered simultaneously at the same time (for example, as two tablets taken together, or as single tablets formulated with each component) or sequentially (for example, one tablet taken after the other). The doses of each component may be taken together (simultaneously) or sequentially, within a few seconds, minutes, days, weeks, or months.

[0102] One component of the combination of the present disclosure may already be being administered to a subject, for example, as part of a standard treatment. In such a case, a second component of the combination of the present disclosure is administered as a second component in treatment to provide the therapeutic combination of the present disclosure.

[0103] Treatment method This disclosure further relates to a method for treating, improving or preventing kidney disease, wherein the method involves a subject a) Therapeutic formulations adapted to deliver approximately 200 mg to 280 mg of repagermanium or propagermanium per day. The present invention provides a method that includes the step of administering a substance.

[0104] This disclosure further relates to a method for treating, improving or preventing kidney disease, wherein the method involves a subject i) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. The present invention provides a method that includes the step of administering a substance.

[0105] This disclosure further relates to a method for treating, improving or preventing kidney disease, wherein the method involves a subject i) A therapeutically effective amount of repagermanium or propagermanium per day; and ii) A therapeutically effective amount of AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone. The present invention provides a method that includes the step of administering a substance.

[0106] This disclosure further relates to a method for treating, improving or preventing kidney disease, wherein the method involves a subject i) Therapeutic formulations adapted to deliver approximately 200 mg to 280 mg of repagermanium or propagermanium per day; and ii) A therapeutically effective dose of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone. The present invention provides a method that includes the step of administering a substance.

[0107] The subjects to be treated may be mammals (including human mammals).

[0108] The above-mentioned repagermanium and the above-mentioned AT1R blocker may be administered in the same dosage form or in separate dosage forms.

[0109] The repagermanium and / or AT1R blocker described above may be antibody inhibitors or blockers of their respective receptors. The repagermanium and / or AT1R blocker described above may be the same active agent, for example, a bispecific antibody. The repagermanium and / or AT1R blocker described above may be a pharmaceutically acceptable salt of the repagermanium and / or AT1R blocker described above.

[0110] The above-mentioned repagermanium and the above-mentioned AT1R blocker may be administered simultaneously or sequentially.

[0111] One component of the combination of the present disclosure may already be being administered to a subject, for example, as part of a standard treatment. In such a case, a second component of the combination of the present disclosure is administered as a second component in treatment to provide the therapeutic combination of the present disclosure.

[0112] While not intended to be limited to any particular mode of action, in one embodiment, the repagermanium has a higher affinity and / or potency and / or efficacy when the CCR2 interacts with or modulates a downstream pathway of the CCR2 when the CCR2 is associated with the angiotensin receptor AT1R. For example, the CCR2 and the angiotensin receptor AT1R may associate as a CCR2 / AT1R complex.

[0113] Where necessary, the combinations described herein provide lower doses of either the AT1R blocker or repagermanium than those obtained when either one is used alone. For example, either or both of the AT1R blocker or repagermanium may be provided in sub-therapeutic doses. This has the benefit of reducing the negative safety profile of either the AT1R or repagermanium while maintaining the same therapeutic benefit.

[0114] In further embodiments, when the repagermanium is administered to a subject simultaneously with or sequentially to an AT1R blocker, the affinity, potency, and / or efficacy of the combination are greater than those achieved when the repagermanium is not administered in combination with the AT1R blocker (whether simultaneously or sequentially). In further embodiments, the synergistic effect (as measured by affinity, potency, and / or efficacy) is achieved when the repagermanium is administered to a subject in combination with the AT1R blocker (whether simultaneously or sequentially).

[0115] While not intended to be limited to any particular mode of action, in one embodiment, the AT1R blocker has a higher affinity and / or potency and / or efficacy when it interacts with the angiotensin receptor AT1R when the angiotensin receptor AT1R is associated with CCR2. For example, the CCR2 and the angiotensin receptor AT1R may associate as a CCR2 / AT1R complex. In a further embodiment, when the AT1R blocker is administered to a subject simultaneously with or sequentially with repagermanium, the affinity, potency and / or efficacy of the combination is greater than the affinity, potency and / or efficacy achieved when the AT1R blocker is not administered in combination with repagermanium (whether simultaneously or sequentially). In further embodiments, synergistic effects (as measured by affinity, potency, and / or efficacy) are achieved when the AT1R blocker is administered to the subject in combination with repagermanium (whether simultaneously or sequentially).

[0116] For the purposes of this disclosure, the same or higher affinity of an AT1R blocker to the CCR2-AT1R complex is considered to be AT1R blocker affinity when there is a loss of 3.5 times or less in affinity to the CCR2-AT1R complex compared to affinity to AT1R.

[0117] delivery The dosage forms provided by this disclosure may further include vials, cartridges, containers, tablets, or capsules containing the pharmaceutical formulations of this disclosure, together with instructions for administering the dosage forms to a subject for the treatment, improvement, or prevention of a condition or disease.

[0118] The amount of each active ingredient that can be combined with a carrier substance to produce a single dose varies depending on the host to be treated and the specific mode of administration. For example, a formulation intended for oral administration to humans may contain approximately 0.5 mg to 1 g of each active compound, along with an appropriate and convenient amount of carrier substance (which can vary from approximately 5 to 95% of the total formulation). Dosage units generally contain between approximately 0.5 mg and 500 mg of the active ingredient.

[0119] If necessary, the above AT1R blockers are supplied in doses of approximately 2 mg to 800 mg / day, and are supplied in doses of 1 or more. For example, the above AT1R blockers may be supplied in doses of approximately 75 mg to 300 mg / day. For example, the above AT1R blocker is irbesartan, administered in doses of approximately 75 mg, 150 mg, or 300 mg per day, and is supplied in doses of 1 or more.

[0120] If necessary, the above-mentioned repagermanium is supplied in doses of approximately 200 mg to 280 mg of repagermanium or propagermanium per day, and is supplied in doses of 1 or more. For example, the above-mentioned repagermanium or propagermanium is supplied in doses of approximately 220 mg to 260 mg / day, or approximately 240 mg / day, and is supplied in doses of 1 or more.

[0121] The dose of each active agent may be provided in a single dosage form or in two separate dosage forms, and may include approximately 2 mg to 800 mg of the AT1R blocker and approximately 200 mg to 280 mg of the repagermanium or propagermanium. The dose of the two active agents may be provided in a single dosage form or in two separate dosage forms, and may include (i) a daily dose of approximately 2 mg to 800 mg of the AT1R blocker and (ii) a daily dose of approximately 200 mg to 280 mg of the repagermanium or propagermanium. The AT1R blocker may be irbesartan, and the dosage form may include a daily dose of approximately 300 mg of irbesartan. Alternatively, the AT1R blocker may be valsartan, administered in doses of approximately 80 mg, 160 mg, or 320 mg per day, and provided in one or more doses.

[0122] However, it is understood that a particular dose level for any given subject depends on various factors, including the activity of the specific compound used, age, weight, overall health, sex, diet, administration time, route of administration, elimination rate, drug combination, and the severity of the particular condition or disease being treated.

[0123] The formulations of this disclosure may be administered by injection in various situations, or may be prepared for oral, pulmonary, nasal or any other form of administration. The formulations may be administered, for example, intravenously, subcutaneously, intramuscularly, intraorbitally, ophthalmologically, intraventricularly, intracranially, intrasacrally, intraspinally, intracisionally, intracisionally, intraperitoneally, orally, rectally, vaginally, nasally, or by aerosol administration.

[0124] The mode of administration is among at least one aspect suitable for the form in which the above-mentioned formulation is prepared. The mode of administration for the most effective response may be determined empirically, and the means of administration described below are shown as examples and are not in any way limiting the methods of delivery of the formulations of this disclosure. All formulations provided are commonly used in the pharmaceutical industry and are generally known to appropriately qualified physicians.

[0125] Oral administration form Oral solid-state formulations are intended for use herein and are generally described in Chapter 89 of Martin, Remington's Pharmaceutical Sciences, 18th edition (1990 Mack Publishing Co. Easton PA 18042), which is incorporated herein by reference. Examples of solid-state formulations include tablets, capsules, pills, lozenges, cachets, or pellets. Liposome or proteinoid encapsulation may also be used to formulate the formulations of the present invention (e.g., as proteinoid microspheres reported in U.S. Patent No. 4,925,673). Liposome encapsulation may be used, and the liposomes may be derivatized with various polymers (e.g., U.S. Patent No. 5,013,556). Possible solid-state formulations for the above therapeutic agents are described in Marshall, in Modern Pharmaceutics, Chapter 10, edited by Banker and Rhodes, (1979) (as incorporated herein by reference). Generally, the formulations include the compounds described as part of this disclosure (or chemically modified forms thereof) and an inert component that provides protection to the gastric environment and allows for the release of biologically active substances in the intestine.

[0126] The oral administration forms of this disclosure may be liquids, suspensions, or other suitable administration forms delivered directly into a feeding tube of a patient who is in a comatose state and / or connected to a ventilator and is unable to swallow tablets or the like.

[0127] With respect to the repagermanium or AT1R blockers of this disclosure, the site of release may be the stomach, small intestine (duodenum, jejunum, or ileum), or large intestine. Those skilled in the art have available formulations that do not dissolve in the stomach but release the substance in the duodenum or other part of the intestine. In one aspect, the release avoids adverse effects on the gastric environment by either protecting the formulation or releasing the compound beyond the gastric environment, for example, in the intestine.

[0128] Coatings or mixtures of coatings may also be used for tablets (which are not intended to protect against stomach irritation). These include, but are not limited to, sugar coatings or coatings that make tablets easier to swallow. Exemplary capsules consist of a hard shell (e.g., gelatin) for the delivery of dry therapeutic agents (i.e., powders); for liquid forms, a soft gelatin shell may be used. In certain aspects, the shell material for a cachet is thick starch or other edible paper. For pills, lozenges, molded tablets or crushed tablets, moist massing techniques may also be considered, but are not limited to.

[0129] This disclosure further provides therapeutically effective pharmaceutical formulations and orally sustained-release pharmaceutical formulations comprising release delay agents, in accordance with this disclosure.

[0130] In one aspect of this disclosure, the release retarder is a water-soluble, water-swellable, and / or water-insoluble polymer. In particular, the water-soluble polymer is selected from the group including ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, enteric coatings, and semipermeable membranes. In another aspect of this disclosure, the release retarder is a nonpolymeric release retarder. More specifically, the nonpolymeric release retarder is hydrogenated castor oil. The formulations of this disclosure may be milled or granulated, compressed into tablets, or encapsulated in capsules according to conventional procedures known in the art.

[0131] To ensure sufficient gastric tolerance, an impermeable coating to at least pH 5.0 is used. Examples of more common inert ingredients used as enteric coatings include cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and Shellac. These coatings can be used as mixed films.

[0132] As used herein, the term “sustained release” means the gradual but continuous release or sustained release of the therapeutic compound contents over a relatively long period after oral ingestion. The release may continue after the pharmaceutical preparation has passed through the stomach and until it reaches and after it reaches the intestines. The term “sustained release” also means delayed release, in which the release of the therapeutic compound is delayed for a period of time, for example, until the pharmaceutical preparation reaches the intestines, rather than starting immediately after the pharmaceutical preparation reaches the stomach. Once it reaches the intestines, an increase in pH may induce the release of the therapeutic compound from the pharmaceutical preparation.

[0133] The term "release retardant," as used herein, means a substance that reduces the rate of release of a therapeutic compound from a pharmaceutical formulation when taken orally. The release retardant may be polymer or nonpolymer. The release retardant may be used according to one of several sustained-release systems, including, for example, a diffusion system, a dissolution system, and / or an osmotic system.

[0134] In certain contexts, the therapeutic agent is contained in the formulation as a multitude of fine particles in the form of granules or pellets with a particle size of approximately 1 mm. Formulations of the substance for capsule administration are, in certain contexts, powders, lightly compressed plugs, or even tablets. In one context, the therapeutic agent may be prepared by compression.

[0135] Colorants and flavoring agents may be included as needed. For example, the compound may be formulated (e.g., liposome or microsphere encapsulation) and then further incorporated into an edible product (e.g., a refrigerated beverage containing colorants and flavoring agents).

[0136] The volume of the above-mentioned therapeutic agent may, in one instance, be diluted or increased with an inert substance. These diluents may include carbohydrates, particularly mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextran, and starch. Certain inorganic salts may also be used as fillers (including tricalcium triphosphate, magnesium carbonate, and sodium chloride) as needed. Some commercially available diluents include Fast-Flo, Emdex, STA-Rx 1500, Emcompress, and Avicell.

[0137] In other embodiments, the disintegrant is included in the formulation of the therapeutic agent in a solid dosage form. Substances used as disintegrants include, but are not limited to, commercially available starch-based disintegrants, starch containing Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultraamylopectin, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, natural sponge, and bentonite are also intended. Another form of the disintegrant is an insoluble cation exchange resin. Powdered gums are also used as disintegrants and binders as needed, and these include, but are not limited to, powdered gums (e.g., agar, karaya, or tragacanth). Alginic acid and its sodium salts are also useful as disintegrants.

[0138] The binder is intended to hold the therapeutic compounds together to form hard tablets and is a substance derived from natural products, such as acacia, tragacanth, starch, and gelatin, but is not limited to these. Other binders include, but are not limited to, methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropyl methylcellulose (HPMC) are intended for use in alcoholic solutions for granulating the therapeutic agents.

[0139] Antifrictional agents may be included in the formulation of the therapeutic agent as needed to prevent adhesion during the formulation process. Lubricants may be used as a layer between the therapeutic agent and the die wall as needed, and these may include, but are not limited to, stearic acid (including its magnesium and calcium salts), polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Exemplary soluble lubricants may also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, and Carbowax 4000 and 6000.

[0140] A glidant may be added as needed to improve the flow properties of the above compound during formulation and to assist in rearrangement during compression. Examples of such glidants include, but are not limited to, starch, talc, calcined silica, and hydrated silicoaluminate.

[0141] To aid in the dissolution of the above therapeutic agents in an aqueous environment, surfactants may be added as wetting agents in certain embodiments. Examples of surfactants, but not limited to them, include anionic detergents (e.g., sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and sodium dioctyl sulfonate). Cationic detergents may be used as needed, and may include, but are not limited to, benzalkonium chloride or benzethonium chloride. A list of potential nonionic detergents that may be included in the above formulations as surfactants includes lauromacrogol 400, polyoxy 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. When used, these surfactants may be present in the formulations of the above compounds either alone or in mixtures of different proportions.

[0142] Additives that can potentially enhance the uptake of the above compounds may be desirable. Examples of such additives include the fatty acids oleic acid, linoleic acid, and linolenic acid.

[0143] Controlled-release formulations may be desirable. In certain contexts, the compound can be incorporated into an inert matrix (i.e., gum) that enables release by either diffusion or leakage mechanisms. In some contexts, a slowly denaturing matrix can also be incorporated into the formulation. Another form of controlled release of this therapeutic agent is OROS. TM The method is based on the Therapeutic Drug System (Alza Corp.). Specifically, the drug is encapsulated in a semipermeable membrane that, due to its osmotic effect, allows water to enter and push the drug out through a single small opening. Some enteric coatings may also have a delayed-release effect.

[0144] In other contexts, mixtures of materials may be used to provide an optimal film coating. Film coating may be carried out, for example but not limited to, in a pan coater, in a fluidized bed, or by compression coating.

[0145] Dosage schedule It is recognized that in certain circumstances, the formulations of this disclosure may be administered as a single-dose schedule or as a multi-dose schedule. A multi-dose schedule may involve a primary delivery process using 1 to 10 distinct doses, with other doses administered at subsequent time intervals as needed to maintain or enhance the treatment. The administration schedule may also be determined, at least in part, by the individual's needs and the physician's judgment.

[0146] This disclosure therefore provides tablets, capsules, liquids, suspensions or other oral administration forms comprising the pharmaceutical preparations of this disclosure.

[0147] The AT1R blocker and the repagermanium or propagermanium may be delivered in the same formulation or in separate formulations.

[0148] The above AT1R blocker and the above repagermanium or propagermanium may be administered in the same form or in different forms.

[0149] A subject receiving the AT1R blocker and the repagermanium may have already received one of the active agents and may be administered other components of the treatment of this disclosure in accordance with this disclosure.

[0150] use This disclosure provides the use of a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium for the manufacture of formulations for the treatment, improvement, or prevention of kidney disease.

[0151] This disclosure provides the use of a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium for the treatment, improvement, or prevention of kidney disease.

[0152] This disclosure provides the use of a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of an AT1R blocker for AT1R alone, for the manufacture of formulations for the treatment, improvement, or prevention of kidney disease.

[0153] This disclosure provides the use of a therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of an AT1R blocker for AT1R alone, for the treatment, improvement, or prevention of kidney disease.

[0154] The disclosure also provides for the use of (a) a therapeutically effective amount of repagermanium or propagermanium, and (b) a therapeutically effective amount of an AT1R blocker having the same or greater affinity for the CCR2-AT1R complex as the affinity for AT1R alone, for the manufacture of formulations for the treatment, improvement or prevention of kidney disease.

[0155] The disclosure also provides for the use of (a) a therapeutically effective amount of repagermanium or propagermanium, and (b) a therapeutically effective amount of an AT1R blocker having the same or greater affinity for the CCR2-AT1R complex as the affinity for AT1R alone, for the treatment, improvement or prevention of kidney disease.

[0156] The disclosure further provides therapeutically effective amounts of repagermanium or propagermanium for use in formulations for the treatment, improvement or prevention of kidney disease, wherein at least one AT1R blocker is administered to the subject simultaneously with or sequentially with the repagermanium or propagermanium.

[0157] The Disclosure further provides therapeutically effective amounts of repagermanium or propagermanium for use in formulations for the treatment, improvement or prevention of kidney disease, wherein the repagermanium is administered to a subject simultaneously with or sequentially with a therapeutically effective amount of AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of an AT1R blocker for AT1R alone.

[0158] The therapeutically effective dose of repagermanium or propagermanium described above may be approximately 200 mg to 280 mg of repagermanium or propagermanium per day.

[0159] The AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone may be selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan. In one embodiment, the AT1R blocker is not EXP3174.

[0160] The therapeutically effective dose of the above-mentioned AT1R blockers may be approximately 2 mg to 800 mg per day.

[0161] The above-mentioned repagermanium or propagermanium and the above-mentioned AT1R blocker may be administered in the same dosage form or in separate dosage forms.

[0162] The above-mentioned repagermanium or propagermanium and / or AT1R blocker may be antibody inhibitors or blockers of their respective receptors. The above-mentioned repagermanium or propagermanium and the AT1R blocker may be the same active agent, for example, a bispecific antibody. The above-mentioned repagermanium or propagermanium and / or AT1R blocker may be a pharmaceutically acceptable salt of the above-mentioned repagermanium or propagermanium and / or AT1R blocker.

[0163] The above-mentioned repagermanium or propagermanium and the above-mentioned AT1R blocker may be administered simultaneously or sequentially.

[0164] kit This disclosure relates to a kit for the treatment or prevention of kidney disease, the kit being: a) A daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium; and b) Provide a kit including instructions for use.

[0165] This disclosure further relates to a kit for the treatment or prevention of kidney disease, the kit comprising: a) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity of the AT1R blocker for AT1R alone; b) Instructions for use, We provide a kit that includes this.

[0166] This disclosure relates to a kit for the treatment or prevention of kidney disease, the kit being: a) A therapeutically effective amount of repagermanium or propagermanium; b) A therapeutically effective amount of an AT1R blocker having an affinity for the CCR2-AT1R complex equal to or greater than the affinity for AT1R alone; and c) Instructions for use, We provide a kit that includes this.

[0167] The therapeutically effective dose of repagermanium or propagermanium described above may be approximately 200 mg to 280 mg of repagermanium or propagermanium per day.

[0168] The AT1R blocker having an affinity for the CCR2-AT1R complex that is equal to or greater than the affinity of the AT1R blocker for AT1R alone may be selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, and spalsentan. In one embodiment, the AT1R blocker is not EXP3174.

[0169] The contents of the above kit may be freeze-dried, and the kit may further contain a solvent suitable for the reconstruction of its freeze-dried components. Each component of the above kit may be packaged in a separate container and associated with such container may be a warning in the form prescribed by a government agency that regulates the manufacture, use, or sale of a pharmaceutical or biological product, and such warning may reflect the approval by the regulatory authority for manufacture, use, or sale of human administration.

[0170] If the components of the above kit are provided in one or more liquid solutions, the liquid solutions may be aqueous solutions, for example, sterile aqueous solutions. For in vivo use, the expression construct may be formulated into a pharmaceutically acceptable syringe. In this case, the container means may itself be an inhalant, syringe, pipette, eye dropper, or other such similar device. From these, the formulation may be applied to the affected area of ​​an animal (e.g., the lungs), injected into the animal, or further applied to and mixed with other components of the above kit.

[0171] The components of the above kit may also be supplied in a dry or lyophilized form. If the reagents or components are supplied in a dry form, reconstitution is generally by the addition of a suitable solvent. It is assumed that the solvent may also be supplied in a separate container. Regardless of the number or type of containers, the kit of this disclosure may also include, or be packaged with, instructions for assisting in the injection / administration or placement of the final combined formulation into the body of an animal. Such instructions may be inhalants, syringes, pipettes, forceps, measuring spoons, eyedroppers, or any such medically approved delivery vehicle.

[0172] general Those skilled in the art will recognize that the inventions described herein are subject to variations and modifications other than those specifically described. The present invention encompasses all such variations and modifications. The present invention also encompasses all of the steps, features, formulations and compounds mentioned or shown herein, individually or collectively, as well as any and all combinations of any two or more of the above steps or features.

[0173] Each document, reference, patent application, or patent cited herein is expressly incorporated herein by reference in its entirety, meaning that the reader should read and consider them as part of the text. The absence of repetition of any document, reference, patent application, or patent cited herein is solely for the sake of brevity.

[0174] Descriptions, statements, product specifications, and product sheets of any manufacturer of any product in any document mentioned herein or incorporated herein by reference are incorporated herein by reference and may be used in the practice of the present invention.

[0175] The present invention should not be limited in scope by any of the specific embodiments described herein. These embodiments are intended to be illustrative only. Functionally equivalent products, formulations, and methods clearly fall within the scope of the invention as described herein.

[0176] The inventions described herein may include one or more ranges of values ​​(e.g., size, displacement, and electric field strength). A range of values ​​is understood to include all values ​​within that range, including the value that defines the range and the values ​​immediately adjacent to that value that define the boundary of the range, and values ​​adjacent to that range that produce the same or substantially the same result. Thus, unless otherwise indicated, the numerical parameters shown herein and in the claims are approximations that may vary depending on the desired characteristics to be obtained by the invention. Thus, “about 80%” means “about 80%” and also “80%”. At a minimum, each numerical parameter should be interpreted in light of the number of significant figures and common rounding methods.

[0177] Throughout this specification, unless otherwise specified by context, variations of the word “comprise,” “comprises,” or “comprising” are understood to imply the inclusion of the integer or group of integers mentioned, but not the exclusion of any other integer or group of integers. In this disclosure, and in particular in the claims and / or paragraphs, terms such as “comprises,” “comprised,” and “comprising” may have meanings under U.S. patent law; for example, they may mean “includes,” “included,” and “including”; it is also noted that terms such as “consisting essentially of” and “consists essentially of” may have meanings under U.S. patent law, for example, they allow for elements not expressly described but exclude elements found in the prior art or that would affect the fundamental or novel features of the present invention.

[0178] Other definitions of selected terms used herein may be found in the detailed description of the invention and may apply throughout. Unless otherwise defined, all other scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention pertains. The term “active agent” may mean one active agent or may encompass two or more active agents.

[0179] The following embodiments serve to more fully describe ways in which the above invention is used and to illustrate the best modes intended for carrying out various aspects of the invention. It is understood that these methods are not intended in any way to limit the true scope of the invention, but rather are provided for illustrative purposes. [Examples]

[0180] Examples Further features of the present invention are better described in the following non-limiting embodiments. This description is included solely for illustrative purposes of the invention and should not be understood as a limitation to the broader description of the invention as set forth above.

[0181] Example 1 Pharmacokinetic model of repagermanium method Pharmacokinetic data were available from studies DMX-200-101, DMX-200-201, DMX-200-202, and DMX-200-203. Table 2: Pharmacokinetic data [Table 2]

[0182] In all studies, blood samples were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS / MS). This method involves adding deuterated PPG (d2-PPG) as an internal standard (IS) and liquid-liquid extraction with chloroform for sample preparation. The samples were analyzed using an LC-MS / MS system consisting of a Shimadzu LC20AD with a Thermo Fisher Hypercarb column using gradient elution and an API 4000 mass spectrometer. The mass spectrometer was operated in negative electrospray ionization (ESI) ion mode and used to monitor the transition from precursor ions to product ions for PPG and d2-PPG, respectively. The limit of quantification (LLOQ) was 5 ng / mL.

[0183] Pharmacometric analysis was performed in several steps, driven by the suitability of clinical data for the development of the PK model. 1. An initial structured population PK model was developed using data from healthy volunteers (Study 101). 2. This model was subsequently updated using sparse data from patients in Studies 202 and 203, and the effects of covariates were explored. 3. Model quality certification was performed. 4. Finally, the developed model was used to simulate the effects of covariates on key PK parameters and on potential exposures.

[0184] SAS software (version 9.4) was used to prepare the analysis dataset. Population PK analysis was performed using NONMEM (version 7.5.0) and conditional first-order estimation with interactions (FOCE). PsN (version ≥ 5.0.0) was also used to perform NONMEM execution, as well as visual predictive check (VPC), sampling importance resampling (SIR), and automated stepwise covariate modeling (SCM). R software (version ≥ 4.0.2) was used for exploratory analysis, goodness of fit (GOF) plot creation, and summary statistics. Campsis (version 1.1.0), an R-based software application provided by Calvagone, was used for simulations.

[0185] NONMEM version 7.5.0 was used with the GNU Fortran (GCC) compiler version 9.3.0 on a fault-tolerant VMWare ESXi 7.0u1 platform with an Intel Xeon E5-2640 v3 (8 HT 2.60GHz) and a compute node with an Intel Xeon E5-2680 v4 14HT 2.40GHz, running on an Ubuntu 20.04 LTS system with Linux® kernel version 5.4.0-72-generic. The Fortran compiler optimization options were: -static -w -O3 -ffast-math. All software was used in a verified environment.

[0186] In mixed-effects modeling, population parameters are categorized as follows: Fixed-effects parameters, θ (theta), are used for structural population parameters (e.g., Ka, CL, V, F, etc.), as well as for specific parameters that relate the above population parameters to physiological or demographic variables (covariates) included in the analysis. Random-effects parameters, η (eta), are used to describe between-individual variability (IIV) in fixed-effects parameters, and may also be used to describe time-interval variability (IOV) when appropriately included in the above model. The η parameter is assumed to be a normally distributed random variable with a mean of zero and a variance-covariance matrix Ω (omega). The square roots of the diagonal elements of this matrix provide the IIV estimates. Elements outside the diagonals (if not fixed to zero) are generally translated into correlations. Random-effects parameters, ε (epsilon), are used to describe residual variability. Residual variability includes inter-individual errors, analytical errors, potential errors due to model misspecification, and any other errors not accounted for by structural elements such as θ and η. The residual variability parameter is assumed to be a normally distributed random variable with mean zero and standard deviation σ (sigma).

[0187] Different residual error models, and, where necessary, different structural models combined with different estimation methods, were applied to identify the most appropriate mixed-effects model. Parameter estimation was performed using conditional first-order estimation (FOCE) with an interaction option to account for the interaction between η (inter-individual random effect) and ε (residual random effect). An exponential model was used to describe the inter-individual variability (IIV) of the model parameters, and a log-normal distribution was assumed for the individual pharmacokinetic parameters:

number

[0188] Covariate model construction is exploratory. Initially, potential relationships between covariates and individual parameter estimates are explored on a graph, assuming a reasonably low degree of eta contraction in the parameters (<25%). Since the model is intended for use in child exposure simulations, the relationship between body weight and PK parameters was decided to be considered according to allometric theory, as already done in the base model. A fixed allometric exponent of 0.75 was used for clearance and inter-compartment flow (Q), while an exponent of 1.0 was used for the relationship between volume and body weight. Automated stepwise covariate modeling (SCM) using forward add / backward delete procedures, as implemented in PsN

[20] , was applied to the base model on the fully analyzed dataset. To improve the computation time of the above SCM, PsN generates a linearized form of the model to which the Taylor transform is applied, and then the covariate functions are applied to it. The final model from the SCM procedure was then re-run in its original untransformed form. The following covariates were tested in the above model for their association with CL and relative bioavailability (F): weight, BMI, age, race (if non-white participants exceeded 5%), sex, liver / renal function (AST, ALT, albumin, CRCL, eGFR), and disease (equal to the study). In principle, baseline covariate values ​​were considered, but an assessment was made as to whether changes in covariates over time should also be considered in the covariate model. Correlations between covariates were evaluated graphically. If two or more covariates were highly correlated, the most clinically / biologically and practically relevant covariate was prioritized for testing in the covariate analysis, and other covariates were discarded or tested only after more relevant ones had been included in the above model. For continuous covariates, a linear or power function was tested, as shown in the following formula:

number

number

[0189] result The clinical effect of reducing the protein-to-creatinine ratio, a biomarker of renal function, is induced by administration of a 120 mg BID dose of repagermanium, which results in a steady state of approximately 15 μg.h / mL (range 5-25) AUC, as shown in Table 3. Table 3: Individual steady-state exposure (AUC) for the total daily dose of 240 mg in subjects evaluated in the analysis. 24hr Summary statistics of ) [Table 3]

[0190] This specific dose and exposure of repagermanium is required for peak renal efficacy.

[0191] Example 2 The above AT 1 ARB efficacy in the R-CCR2 complex is AT 1 ARB-CCR2 inhibitor combinations that inhibit G protein activation are not less effective than the predicted efficacy of ARBs against R alone for the treatment of diseases. The CCR2-AT1R complex has been previously identified as a target for the treatment of kidney disease. Inhibition of the CCR2 pathway and AT1R signaling is required for clinical efficacy.

[0192] The inventors previously demonstrated that the above AT1R / CCR2 heteromer constitutively inhibits inositol phosphate signaling with ARBs. The inventors also showed that the above CCR2-AT1R complex results in the activation of Gαi proteins and the recruitment of β-arrestin.

[0193] However, it was unknown whether ARBs, when acting on the CCR2-AT1R complex, maintain their efficacy in inhibiting Gαi protein activation or mobilizing β-arrestin compared to AT1R alone. Understanding the ability of ARBs to inhibit AT1R in the aforementioned CCR2-AT1R complex is important because it is expected to affect clinical efficacy.

[0194] For example, we have shown that AngII inhibits Gαi1-coupled MCP-1-inducible activation by the CCR2-AT1R complex. Blocking this AngII-mediated response using an ARB is expected to eliminate this inhibition of MCP-1-inducible Gαi1 signaling.

[0195] This experiment evaluated the efficacy of ARBs in AT1R alone compared to the efficacy of ARBs in the co-stimulated CCR2-AT1R complex in a Gαi protein dissociation assay.

[0196] method To establish whether ARBs maintain their efficacy in inhibiting Gαi protein activation, the efficacy of ARBs in AT1R alone correlated with the efficacy of ARBs in the co-stimulated CCR2-AT1R complex obtained from a Gαi protein dissociation assay (Figure 2).

[0197] The efficacy of ARBs in AT1R alone was obtained from the IUPHAR pharmacological guide database and / or publicly available sources such as Michel et al. (2013). The efficacy of ARBs in the co-stimulated CCR2-AT1R complex described above was experimentally derived as outlined below.

[0198] HEK293FT cells were transiently transfected with plasmids encoding Gαi1-Nluc and CCR2-YFP hemagglutinin (HA)-tagged AT1R. 48 hours after transfection, cells were incubated with substrate for 1 hour, and then pre-incubated for 30 minutes with various concentrations of ARBs (candesartan cilexetil, losartan, telmisartan, irbesartan, eprosartan, EXP3174, valsartan, olmesartan medoxomil, olmesartan, azilsartan camedoxomil, or candesartan). Cells were then treated with 100 nM AngII and 100 nM MCP-1 or vehicle, and ligand-inducible BRET was measured by measuring continuous light emission at 400–475 nm and 520–540 nm. The above BRET signal was calculated by subtracting the ratio of 520-540 nm emission to 400-475 nm emission for vehicle-treated cell samples from the same ratio for a second aliquot of the same cells treated with the ligand (ligand-induced BRET).

[0199] Next, using GraphPad Prism (v9.3.1), we plotted ligand-induced BRET against ARB concentration and measured the potency (pIC). 50 The -value was obtained by fitting a sigmoid concentration response curve using the nonlinear regression function of GraphPad Prism (v9.3.1).

[0200] To calculate the potency of spalsentan, HEK293FT cells were transiently transfected with plasmids encoding Gαi1-Rluc8, CCR2-YFP, and hemagglutinin (HA)-tagged AT1R. 48 hours after transfection, the cells were incubated with substrate for 2 hours, then pre-incubated with spalsentan (1 μM), and filtered light emission was measured continuously for 30 minutes at 1 second each in the "donor wavelength window" (410–490 nm) and the "acceptor wavelength window" (520–620 nm). Subsequently, the cells were stimulated with 10 nM MCP-1 (CCL2) and various concentrations of AngII, and filtered light emission was measured for a further 1 hour.

[0201] Next, using GraphPad Prism (v9.3.1), ligand-induced BRET was plotted against AngII concentration in the absence and presence of sparnentan (1 μM). Sigmoid concentration response curves were fitted to the data using the nonlinear regression function of GraphPad Prism (v9.3.1), and sparnentan affinity estimates (pKB) were calculated as pKB, as described by Furchgott ((1972) The classification of adrenoceptors (adrenergic receptors). An evaluation from the standpoint of receptor theory, in Catecholamines, Handbook of Experimental Pharmacology (edited by Blaschko H and Muscholl E) Vol 33, pp 283-335).

[0202] Next, the efficacy of ARBs in the co-stimulated CCR2-AT1R complex was compared to the efficacy of ARBs in AT1R using a linear regression function in GraphPad Prism (v9.3.1). Then, the data in Figure 2 were analyzed using robust linear regression and an outlier removal (ROUT) coefficient set to Q=1 to identify outliers with lower-than-expected efficacy.

[0203] ARBs whose efficacy in the CCR2-AT1R complex was less than one-third of that predicted compared to AT1R alone were considered pharmacologically related, and their statistical significance was evaluated by a one-sample t-test (Figure 3).

[0204] Figure 4 shows the effect of ARB and CCR2 pathway inhibitor combinations on Gαi1 coupling to the CCR2-AT1R complex as measured by ligand-inducible BRET. HEK293FT cells were transiently transfected with plasmids encoding Gαi1-Rluc8 and CCR2-YFP in the presence of hemagglutinin (HA)-tagged AT1R. 48 hours after transfection, cells were used to generate agonist-inducible BRET signal data in live cells stimulated with 100 nM MCP-1 (CCL2) or 100 nM MCP-1 (CCL2) and 100 nM AngII in the presence of irbesartan, a CCR2 pathway inhibitor, or irbesartan and a CCR2 pathway inhibitor.

[0205] During BRET substrate addition, BRET detection was performed in living cells by measuring continuous light emission at 400–475 nm and 520–540 nm before and after agonist addition. The BRET signal was calculated by subtracting the ratio of 520–540 nm emission to 400–475 nm emission for vehicle-treated cell samples from the same ratio for a second aliquot of the same cells treated with the ligand (ligand-induced BRET). CCR2 activation resulted in a decrease in the BRET ratio, indicating that activation caused a conformational change between CCR2 and the pre-assembled Gαi1 protein. Therefore, the above data are shown as "ligand-induced decrease in BRET (percentage of control)," where a change in the BRET signal observed with 100 nM MCP-1 is shown as 100%, and no change in the BRET signal is shown as 0%.

[0206] result The potency values ​​for AT1R alone were obtained from the IUPHAR Pharmacology Guide, Alexander et al, (2021) The Concise Guide to Pharmacology 2021 / 22 Br J Pharmacol. 2021. 178 Suppl 1:S1-S513 and / or Michel et al (2013) A Systematic Comparison of the Properties of Clinically Used Angiotensin II Type 1 Receptor Antagonists Pharmacol Rev 65:809-848. These values ​​represent the predicted potency of ARBs in the co-stimulated CCR2-AT1R complex. Experimental potency values ​​are reported as pIC50 or pKB values. The pIC50 value is the negative logarithm of the molar concentration of ARB required to inhibit the BRET change mediated by the co-stimulated CCR2-AT1R complex by 50%. The experimental potency value reported as pKB is the negative logarithm of the molar concentration that occupies 50% of the receptor at equilibrium in the absence of the agonist.

[0207] A comparison of ARB resistance in AT1R alone and the co-stimulated CCR2-AT1R complex (Figure 2) and Table 4 shows a good linear regression R, indicating a good correlation between the efficacy of ARBs in the co-stimulated CCR2-AT1R complex and AT1R alone. 2 = 0.97 was found.

[0208] This data indicates that the efficacy of most ARBs acting on the CCR2-AT1R complex is within 3.5 times that of ARBs acting on the AT1R receptor alone. This supports the use of ARBs without reduced efficacy when targeting the CCR2-AT1R complex for disease treatment.

[0209] In contrast, outlier analysis identified that EXP3174 had a lower-than-predicted potency in inhibiting co-stimulated CCR2-AT1R complex-mediated Gαi protein activation compared to EXP3147's potency in AT1R alone.

[0210] The efficacy of EX3147 at the CCR2-AT1R complex was 1 / 4.07 compared to that at AT1R alone, which was statistically different (Figure 3; p<0.01, one-sample t-test). This data indicates that the efficacy of ARBs acting at the CCR2-AT1R complex is unclear and cannot be assumed to be the same as that at AT1R alone. ARBs such as EXP3174, which have reduced efficacy at the CCR2-AT1R complex, are undesirable for targeting the CCR2-AT1R complex for disease treatment and are expected to have reduced efficacy.

[0211] Figure 4 shows the changes in ligand-induced BRET after stimulation of cells expressing the CCR2-AT1R complex with vehicle, CCL2 (100 nM), or CCL2 (100 nM) and AngII (100 nM). Gαi1 protein activation in CCR2-AT1R-expressing cells co-stimulated with CCL2 (100 nM) and AngII (100 nM) is inhibited by a combination of ARBs and CCR2 pathway inhibitors. Compared to AT1R alone, the loss of ARB efficacy in the CCR2-AT1R complex is recognized to affect the effectiveness of inhibition.

[0212] Figure 5 and Table 4 show the recognized efficacy of ARBs in combination with the CCR2 pathway inhibitor repagermanium in inhibiting Gαi1 protein activation in CCR2-AT1R-expressing cells co-stimulated with CCL2 (100 nM) and AngII (100 nM). Table 4: Inhibitory efficacy or affinity (pIC) of ARBs in AT1R alone or co-stimulated AT1R+CCR2 complex 50 / pKb) [Table 4] 1. From the IUPHAR Pharmacological Guide and / or Michel et al. (2013). 2. ARB efficacy in inhibiting Gα-protein dissociation from the CCR2 / Luc-AT1R complex co-stimulated with AngII and CCL2 (pIC 50). 3. Predictive ARB efficacy in inhibiting Gα-protein dissociation from the CCR2 / Luc-AT1R complex co-stimulated with AngII and CCL2 in the presence of repagermanium. ^; Fimasartan data are predictive. $, Data from Choung et al, 2018 Discovery of the bifunctional modulator of angiotensin II type 1 receptor (AT1R) and PPARγ derived from the AT1R antagonist, Fimasartan. Bioorganic & Medicinal Chemistry Letters (28), 19:3155-3160. # Spalsentan affinity (pKb) in the AT1R-CCR2 complex. **, p<0.01 One-sample t-test, significant decrease in efficacy compared to AT1R alone.

Claims

1. A therapeutic formulation adapted to deliver a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

2. A therapeutically effective amount of AT １ A therapeutic formulation adapted to deliver a daily dose of an R-blocker, wherein the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ A therapeutic formulation having affinity for the R complex.

3. a) A therapeutically effective amount of repagermanium or propagermanium; and b) A therapeutically effective amount of AT １ An R blocker, wherein the AT １ The AT for only R １ An R blocker having an affinity for the CCR2-AT １ R complex that is the same as or greater than the affinity for the AT １ R blocker, A therapeutic formulation adapted to deliver the daily dose.

4. The formulation according to claim 3, wherein the daily dose of repagermanium or propagermanium is approximately 200 mg to 280 mg.

5. Said AT １ The formulation according to claim 2 or 3, wherein the R-blocker is selected from the list consisting of candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, fimasartan, and spalsentan.

6. Said AT １ The formulation according to claim 2 or 3, wherein the daily dose of the R-blocker is 2 mg to 800 mg.

7. A dosage regimen that includes the step of administering a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium.

8. A therapeutically effective amount of AT １ An administration regimen comprising the step of administering a daily dose of an R-blocker, wherein the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ A dosage regimen that has affinity for the R complex.

9. a) A therapeutically effective amount of repagermanium or propagermanium; and b) A therapeutically effective amount of AT １ It is an R-blocker, １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ AT has affinity for the R complex. １ R-blockers, A dosing regimen that includes the step of administering a daily dose.

10. A method for treating, improving or preventing kidney disease, wherein the method involves a subject a) Therapeutic formulations adapted to deliver approximately 200 mg to 280 mg of repagermanium or propagermanium per day. A method comprising the step of administering a substance.

11. A method for treating, improving or preventing kidney disease, wherein the method involves a subject a) A therapeutically effective amount of AT １ It is an R-blocker, １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ AT has affinity for the R complex. １ R-blockers, A method comprising the step of administering a substance.

12. A method for treating, improving or preventing kidney disease, wherein the method involves a subject a) A therapeutically effective amount of repagermanium or propagermanium per day; and b) A therapeutically effective amount of AT １ It is an R-blocker, １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ AT has affinity for the R complex. １ R-blockers, A method comprising the step of administering a substance.

13. Use of approximately 200 mg to 280 mg of repagermanium or propagermanium per day for the manufacture of formulations for the treatment, improvement, or prevention of renal disease in subjects.

14. Use of approximately 200 mg to 280 mg of repagermanium or propagermanium per day for the treatment, improvement, or prevention of renal disease in subjects.

15. A therapeutically effective amount of AT for the manufacture of formulations for the treatment, improvement, or prevention of kidney disease in subjects. １ The use of an R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ Use that has affinity for the R complex.

16. A therapeutically effective dose of AT for the treatment, improvement, or prevention of kidney disease in a subject. １ The use of an R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ Use that has affinity for the R complex.

17. For the manufacture of a formulation for the treatment, improvement or prevention of kidney disease in a subject, (a) a therapeutically effective amount of repagermanium or propagermanium, and (b) a therapeutically effective amount of AT １ The use of an R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ Use that has affinity for the R complex.

18. For the treatment, improvement, or prevention of kidney disease in subjects, (a) a therapeutically effective amount of repagermanium or propagermanium, and (b) a therapeutically effective amount of AT １ The use of an R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ Use that has affinity for the R complex.

19. A therapeutically effective amount of repagermanium or propagermanium for use in formulations for the treatment, improvement or prevention of kidney disease in a subject, wherein the repagermanium is a therapeutically effective amount of AT １ The R-blocker is administered to the subject simultaneously or sequentially, and the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ A therapeutically effective amount of repagermanium or propagermanium having affinity for the R complex.

20. A therapeutically effective amount of AT for use in formulations for the treatment, improvement, or prevention of kidney disease in subjects. １ An R-blocker, where the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ Having affinity for the R complex, at least one of the aforementioned ATs １ The R-blocker is administered to the subject simultaneously with or sequentially with repagermanium or propagermanium in a therapeutically effective amount of AT. １ R-blocker.

21. A kit for the treatment or prevention of kidney disease in a subject, wherein the kit is a) A daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium; and b) Instructions for use, A kit that includes this.

22. A kit for the treatment or prevention of kidney disease in a subject, wherein the kit is a) A therapeutically effective amount of AT １ An R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ AT has affinity for the R complex. １ R-blockers; b) Instructions for use, A kit that includes this.

23. A kit for the treatment or prevention of kidney disease in a subject, wherein the kit is a) A therapeutically effective amount of repagermanium or propagermanium; b) A therapeutically effective amount of AT １ An R-blocker, the AT １ R-blockers are AT １ The AT for R only １ CCR2-AT with affinity equal to or greater than that of R-blockers １ AT has affinity for the R complex. １ R-blockers; and c) Instructions for use, A kit that includes this.

24. The repagermanium or propagermanium is provided in a daily dose of approximately 200 mg to 280 mg of repagermanium or propagermanium, according to any one of claims 7 to 9, the method according to any one of claims 10 to 12, the use according to any one of claims 13 to 20, or the kit according to any one of claims 21 to 23.

25. Said AT １ The R-blocker is selected from the list consisting of candesartan, eprosartan, fimasartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, fimasartan and spalsentan, according to any one of the administration regimens of claims 7 to 9, the method according to any one of claims 10 to 12, the use according to any one of claims 13 to 20, or the kit according to any one of claims 21 to 23.

26. The subject to be treated is a human mammal, the administration regimen according to any one of claims 7 to 9, the method according to any one of claims 10 to 12, the use according to any one of claims 13 to 20, or the kit according to any one of claims 21 to 23.

27. The aforementioned kidney disease is a kidney disease associated with proteinuria, the administration regimen according to any one of claims 7 to 9, the method according to any one of claims 10 to 12, the use according to any one of claims 13 to 20, or the kit according to any one of claims 21 to 23.

28. The aforementioned kidney disease is selected from a list including focal segmental glomerulosclerosis (FSGS; including idiopathic (primary) FSGS, secondary FSGS, hereditary FSGS, and any other cause of FSGS), chronic kidney disease including chronic kidney disease caused by fibrous disorders of the kidney, immunoglobulin A nephropathy (IgAN), Alport syndrome, and diabetic nephropathy, renal insufficiency (diabetic and non-diabetic), and renal failure conditions including diabetic nephropathy, glomerulonephritis, scleroderma, glomerulosclerosis, proteinuria of primary kidney disease, and renovascular hypertension, and is the administration regimen according to any one of claims 7 to 9, the method according to any one of claims 10 to 12, the use according to any one of claims 13 to 20, or the kit according to any one of claims 21 to 23.

29. Said AT １ The R-blocker is not EXP3174, and the method is one of the administration regimens according to any one of claims 7 to 9, one of the methods according to any one of claims 10 to 12, one of the uses according to any one of claims 13 to 20, or one of the kits according to any one of claims 21 to 23.

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