USE OF A GROUP OF MARKERS FOR DIAGNOSING AND ADJUSTING TREATMENT OF PRIMARY BILIARY CHOLANGITIS, PHARMACEUTICAL COMPOSITION AND SOLID DOSAGE FORM FOR TREATING PRIMARY BILIARY CHOLANGITIS

Abstract

A method of diagnosis and treatment of primary biliary cholangitis is provided. Additionally, a pharmaceutical composition and a solid dosage form for the treatment of primary biliary cholangitis, containing ursodeoxycholic and obeticholic acids, are provided. The technical contribution resides in obtaining a new all-purpose pharmaceutical composition and solid dosage form for the treatment of PBC, which includes both ursodeoxycholic and obeticholic acids, which is effective in use at all stages of PBC and has a complex mechanism of action. In particular, simultaneous blockage of the transport and synthesis of bile acids is achieved.

Claims

1-26. (canceled)

27. A method of diagnosis and treatment of primary biliary cholangitis, wherein it comprises steps of obtaining a patient sample, determining concentration level values of markers IL-6, Nf-kB, MCP-1/CCL2, Bc1-2 in the patient sample, and prescribing to the patient pharmaceutical composition for the treatment of primary biliary cholangitis, wherein prescribing to the patient pharmaceutical composition for the treatment of primary biliary cholangitis is conducted when the markers IL-6, Nf-kB, MCP-1/CCL2, Bc1-2 have the following concentration level values: TABLE-US-00041 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml, and pharmaceutical composition for the treatment of primary biliary cholangitis is the pharmaceutical composition stimulating the FXR nuclear receptor, which leads to inhibiting the bile acid synthesis, as well as achieving anti-inflammatory, anti-fibrotic and anti-apoptotic effects.

28. The method according to claim 27, wherein it comprises determining concentration level values of additional markers TNF-α, NLRP3, MDA, wherein prescribing to the patient pharmaceutical composition for the treatment of primary biliary cholangitis is conducted when the markers IL-6, Nf-kB, MCP-1/CCL2, Bc1-2, TNF-α, NLRP3, MDA, have the following concentration level values: TABLE-US-00042 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml; TNF-α at least 0.14 ng/ml. MDA at least 4.3 nmol/ml.

29. The method according to claim 27, wherein it comprises step of monitoring the patient condition, which comprises such steps as periodically obtaining patient samples, determining concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2 in the patient samples, analyzing the dynamics of the concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2, and correcting patient treatment regimen, wherein correction of the patient treatment regimen is conducted when dynamics of the concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2 does not show an incremental decrease.

30. The method according to claim 29, wherein it comprises determining concentration level values of additional markers TNF-α, NLRP3, MDA, analyzing the dynamics of the concentration level values of the additional markers TNF-α, NLRP3, MDA, and correcting patient treatment regimen, wherein correction of the patient treatment regimen is conducted when dynamics of the concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2, TNF-α, NLRP3, MDA does not show an incremental decrease.

31. A pharmaceutical composition for the treatment of primary biliary cholangitis comprising ursodeoxycholic acid and at least one excipient, wherein it additionally comprises obeticholic acid.

32. The pharmaceutical composition according to claim 31, wherein it is formulated into the dosage form of an uncoated tablet, coated tablet, or capsule.

33. The pharmaceutical composition according to claim 31, wherein it has a modified release of obeticholic acid.

34. The pharmaceutical composition according to claim 31, wherein it has a modified release of ursodeoxycholic acid.

35. The pharmaceutical composition according to claim 31, wherein primary biliary cholangitis in the patient is characterised by the following concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2 in the patient sample: TABLE-US-00043 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml.

36. The pharmaceutical composition according to claim 31, wherein primary biliary cholangitis in the patient is characterised by the following concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2, TNF-α, NLRP3, MDA in the patient sample: TABLE-US-00044 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml; TNF-α at least 0.14 ng/ml; MDA at least 4.3 nmol/ml.

37. A solid dosage form for the treatment of primary biliary cholangitis, wherein it comprises the pharmaceutical composition according to claim 31, wherein the mass ratio of ursodeoxycholic acid to obeticholic acid ranges from 2:1 to 1000:1, and solid dosage form comprises the core comprising ursodeoxycholic acid, and at least one layer comprising obeticholic acid, and is a modified release dosage form.

38. The solid dosage form according to claim 37, wherein the mass ratio of ursodeoxycholic acid to obeticholic acid ranges from 15:1 to 750:1.

39. The solid dosage form according to claim 37, wherein it comprises three layers and the core.

40. The solid dosage form according to claim 39, wherein it comprises an outer layer, a second layer comprising obeticholic acid, a third layer, and a core comprising ursodeoxycholic acid.

41. The solid dosage form according to claim 40, wherein the outer layer is an enteric coating or a gastric acid soluble coating.

42. The solid dosage form according to claim 40, wherein the second layer comprises obeticholic acid in an amount of 1-50 mg.

43. The solid dosage form according to claim 40, wherein the third layer is a STOP layer with a delayed dissolution or an enteric coating.

44. The solid dosage form according to claim 43, wherein the dissolution time of the STOP layer is 1-2 hours.

45. The solid dosage form according to claim 40, wherein the core comprises ursodeoxycholic acid in an amount of 100-1000 mg.

46. The solid dosage form according to claim 40, wherein the core has a modified release of ursodeoxycholic acid within 1-6 hours.

47. The solid dosage form according to claim 40, wherein it comprises the outer layer being the enteric coating, the second layer comprising obeticholic acid, the third layer being STOP layer, and the core comprising ursodeoxycholic acid, wherein the solid dosage form is for the treatment of primary biliary cholangitis which is characterised by the following concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bcl-2 in the patient sample: TABLE-US-00045 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml.

48. The solid dosage form according to claim 47, wherein primary biliary cholangitis is characterised by the following concentration level values of the markers IL-6, Nf-kB, MCP-1/CCL2, Bc1-2, TNF-α, NLRP3, MDA in the patient sample: TABLE-US-00046 IL-6 at least 3.0 pg/ml; Nf-kB at least 14.0 pg/ml; MCP-1/CCL2 at least 215 pg/ml; Bcl-2 at least 0.24 U/ml; TNF-α at least 0.14 hg/ml. MDA at least 4.3 nmol/ml.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0128] FIG. 1 shows a graphical representation of the changes in the marker concentration level values in % of the initial value when using the standard treatment regimen No. 1 (UDCA 15 mg/kg).

[0129] FIG. 2 shows a graphical representation of the changes in the marker concentration level values in % of the initial value when using the provided treatment regimen No. 2 (UDCA 15 mg/kg+OCA 5 mg).

[0130] FIG. 3 shows a graphical representation of the changes in the marker concentration level values in % of the initial value when using the provided treatment regimen No. 3 (UDCA 15 mg/kg+OCA 10 mg).

[0131] FIG. 4 shows the biochemical markers concentration level values.

[0132] FIG. 5 shows the molecular markers concentration level values.

[0133] Since the mechanism of the PBC onset is unknown and it is impossible to clearly diagnose PBC with the definition of the disease stage, the current treatment protocols involve the use of UDCA pharmaceutical compositions, that promote the elimination of FA. Ursodeoxycholic acid is indeed responsible for the transport of FA but does not stop their synthesis. In addition, studies conducted by the authors using the additional group of the markers to determine the processes of inflammation and identify apoptosis and fibrosis in PBC, show that starting from the second stage of PBC, UDCA is no longer effective, since it is already necessary to stop the FA synthesis. Thus, in the absence of the studies on the molecular mechanism and diagnosis of PBC, it was not possible and appropriate to use the pharmaceutical composition that comprises both acids—UDCA and OCA.

[0134] Because compliance is especially important for the PBC patients (as described above), this objective is achieved by developing a solid dosage form for the PBC treatment that comprises the pharmaceutical composition with UDCA and OCA.

[0135] Having conducted their own research, both theoretical and experimental, the authors of the present disclosure concluded that the system for diagnosing and treating PBC, which is used today, has many disadvantages, and needs to be modified.

[0136] Specifically, the use of the new group of the markers for the diagnosis and adjustment of the PBC treatment, that includes the markers, such as IL-6, Nf-kB, MCP-1/CCL2, and Bcl-2, was provided.

[0137] IL-6 (Interleukin 6) is a pro-inflammatory interleukin synthesized by immune cells. It can stimulate the inflammatory process at the site of synthesis. An increase in the amount of IL-6 is observed in the early stages of the disease, when the autoimmune process is just beginning to develop. Its detection in liver biopsy specimens indicates an acute inflammatory process.

[0138] Nf-kB (nuclear factor kappa-light-chain-enhancer of activated B cells) is a protein complex that controls DNA transcription, cytokine synthesis and cell survival. NF-kB is found in almost all types of the animal cells and is involved in cellular responses to stimuli, such as stress, cytokines, free radicals, heavy metals, ultraviolet radiation, lipid peroxidation, and bacterial or viral antigens. NF-kB plays a key role in regulating the immune response. Misregulation of NF-kB is associated with the development of oncology, inflammatory and autoimmune diseases.

[0139] MCP-1/CCL2 (MCP-1, Monocyte chemoattractant protein-1) is one of the key chemokines that regulates the migration and infiltration of monocytes from peripheral blood into tissues. In PBC, it is released by Kupffer cells, and it stimulates the migration of monocytes from the peripheral blood to the liver tissue, which leads to further development of inflammation.

[0140] Bcl-2 is a mitochondrial apoptosis factor. Its elevated value indicates pathological cell death. This pathological phenomenon can be eliminated by neutralizing the primary cause of the cell death (in this case, this is the toxic effect of the bile acids and the autoimmune process).

[0141] In addition to the above markers, TNF-α, NLRP3, and MDA markers can also be studied.

[0142] TNF-α (tumor necrosis factor alpha) is a cytokine, i.e., a small protein used by the immune system for signaling. The primary role of this molecule is the regulation of the activity and metabolism of the immune cells. It can induce inflammation and apoptosis, which may be the cause of many autoimmune diseases. In PBC, an increase in the value of this cytokine is one of the early events; when immune cells are just starting to attack their own cells, its value rises. This happens much earlier than the rise in ALT/AST and bilirubin levels. That is, the detection of this marker in liver biopsy specimens indicates the beginning of the development of the inflammatory process.

[0143] MDA is a marker of inflammation, but from the lipid peroxidation point of view. Its increase indicates a high level of highly reactive oxygen in the tissues, which can be caused by both the inflammation and immune response.

[0144] The new group of the markers is proposed to be used in the diagnosis of PBC in the following way. The patient with suspected PBC is tested for AMA and ANA levels to confirm the presence of the autoimmune processes and establish the PBC. Based on the concentration level values of the claimed group of markers IL-6, Nf-kB, MCP-1/CCL2, and Bcl-2 (potentially additionally TNF-α, NLRP3, and MDA), the stage of PBC is determined. Based on the established stage of PBC, the patient is prescribed specialized treatment. Next, the patient condition is monitored with a possible adjustment of the therapy regimen by the physician.

[0145] As a result of the studies, a scheme for determining the PBC stage was developed based on the concentration level values of the claimed group of the markers. Concentrations of the molecular markers TNF-α, IL-6, Nf-kB, MCP-1/CCL2, Bcl-2 and MDA in the blood of healthy volunteers and patients with PBC at different stages of the disease were studied. Initially, the distribution of patients according to the stages of the disease was made based on the values of generally accepted markers, such as ALT, AST6 GGT and ALP, as well as the results of a clinical examination. Then, venous blood samples were taken from patients at different stages of PBC, and the values of molecular markers TNF-α, IL-6, Nf-kB, MCP-1/CCL2, Bcl-2, and MDA were determined in these samples. Thus, the concentration of molecular markers at different stages of PBC were determined.

[0146] Marker concentration level values are given as a percentage, wherein:

[0147] 0%—the marker is undefinable;

[0148] 100%—the maximum concentration level value of the marker for PBC.

[0149] Stage 1 of PBC is a trigger that triggers damage to liver cells. Reference marker concentration level values:

[0150] 1) IL-6-9.9-10.1%;

[0151] 2) Nf-kB—4.5-5.5%;

[0152] 3) MCP-1/CCL2—4.5-5.5%;

[0153] 4) Bcl-2-4.5-5.5%;

[0154] 5) TNF-α—10.8-13.2%;

[0155] 6) MDA—0-3.3%.

[0156] Stage 2 of PBC—the development of cholestasis and the induction of inflammation. Reference marker concentration level values:

[0157] 1) IL-6-27-33%;

[0158] 2) Nf-kB—31.5-38.5%;

[0159] 3) MCP-1/CCL2—33.3-40.7%;

[0160] 4) Bcl-2-18-22%;

[0161] 5) TNF-α—27.9-34.1%;

[0162] 6) MDA—4.5-5.5%.

[0163] Stage 3 of PBC—the development of high intensity inflammation. Reference marker concentration level values:

[0164] 1) IL-6-72-88%;

[0165] 2) Nf-kB—67.5-82.5%;

[0166] 3) MCP-1/CCL2—7-88%;

[0167] 4) Bcl-2-36-44%;

[0168] 5) TNF-α—58.5-71.5%;

[0169] 6) MDA—22.5-27.5%.

[0170] Stage 4 of PBC—fibrosis. Reference marker concentration level values: 1) IL-6-72-88%;

[0171] 2) Nf-kB—67.5-82.5%;

[0172] 3) MCP-1/CCL2—72-88%;

[0173] 4) Bcl-2-72-88%;

[0174] 5) TNF-α—72.9-89.1%;

[0175] 6) MDA—40.5-49.5%.

[0176] Stage 5 of PBC—cirrhosis. Reference marker concentration level values: 1) IL-6-36-44%;

[0177] 2) Nf-kB—40.5-49.5%;

[0178] 3) MCP-1/CCL2—31.5-38.5%;

[0179] 4) Bcl-2-36-44%;

[0180] 5) TNF-α—45.9-56.1%;

[0181] 6) MDA—58.5-71.5%.

[0182] As can be seen from the scheme above, determining concentration level values of the claimed group of markers enables understanding types and intensity of the pathological processes occurring in the liver and enables clearly differentiating the stages of PBC and adequately prescribing or adjusting the treatment. The claimed markers enable determining PBC already from the first stage of the disease. In addition, the claimed markers demonstrate the occurrence of the inflammation and apoptosis processes already from the initial stages of PBC, thus proving the need for the use of OCA in combination with UDCA starting from the first stages of the disease.

[0183] Moreover, the use of the claimed group of the markers enables diagnosing PBC in the early stage. For example, when the patient has been diagnosed with AMA and ANA antibodies, but liver tests are normal, the physician will most likely not diagnose PBC. As a result, a dangerous situation arises when, without proper treatment, the disease will progress to further stages. At the same time, the study of the claimed group of the markers is more accurate because it will show elevated concentration level values, despite relatively normal liver tests, which determines prescription of drugs.

[0184] PBC Treatment

[0185] The development of PBC is associated with the loss of the immune tolerance to PDC-E2. Defective clearance of apoptotic bodies that are released during the death of the biliary endothelial cells (BECs) is a possible source of autoantigens and further autoimmune reactions. Thus, intact, and antigenically reactive PDC-E2, that was found on the membrane surface of the apoptotic bodies from cultured human cholangiocytes, exhibits on the apotope surface that is absent in other cell types. These apotopes can stimulate the immune response and are available for AMA, which explains the occurrence of the selective autoimmunity against the bile ducts in PBC.

[0186] In addition, the apoptotic bodies released from cholangiocytes, in combination with AMA, stimulate a surge in concentration level values of pro-inflammatory cytokines.

[0187] In addition to PDC-E2, other mitochondrial autoantigens, including the E2 subunit of the oxo-glutarate dehydrogenase complex and the E2 subunit of the branched-chain 2-oxoacid dehydrogenase complex, were also present in the apoptotic bodies that were released from cholangiocytes exposed to the toxic bile acids, and thus an autoimmune reaction occurs.

[0188] During the development of the disease, degrading cells (hepatocytes and cholangiocytes) release mediators that affect sinusoidal cells, among which are stellate cells—the main cells that synthesize the matrix in the damaged liver. In the normal condition, stellate cells are located in the Disse space, and they are the main depot for vitamin A and are involved in the regulation of fibrogenesis. As a result of the progressive chronic injury, stellate cells become activated and differentiate into myofibroblast-like cells, acquiring contractile, proinflammatory, and profibrotic properties. In the future, this leads to a disruption in the interaction of fibrous and antifibrotic mechanisms, which leads to excessive synthesis of the extracellular matrix and the formation of liver fibrosis.

[0189] In particular, Kupffer cells, regional macrophages of the liver, are important effector cells for the inflammation development. When liver cell death starts as a result of the autoimmune processes, it is Kupffer cells that react to damage-associated molecular fragments that are released from dead cells (DAMP) and stimulate further inflammation. It is they that recruit circulating blood monocytes to the liver, and stimulate the stellate cells of the liver, which begin to produce connective tissue, which leads to fibrosis. This, in turn, leads to the further development of the inflammation, fibrosis, cirrhosis and liver cancer.

[0190] As mentioned above, currently for the PBC treatment, pharmaceutical compositions comprising ursodeoxycholic acid are used first, and, in the absence of the effectiveness or in the case of intolerance, pharmaceutical compositions with obeticholic acid are prescribed. In order to better understand the disadvantages of this approach, it is necessary to consider the mechanisms of action of both acids.

[0191] Experimental data indicate three main mechanisms of action of UDCA: [0192] protection of cholangiocytes against cytotoxic hydrophobic bile acids; [0193] stimulation of hepatobiliary secretion; [0194] protection of hepatocytes from apoptosis caused by bile acids.

[0195] One of the protective mechanisms of UDCA against cholestasis is the utilization of the accumulated bile acids. However, the secretory ability of hepatocytes is closely related to the presence of the transport proteins in the tubular membrane. In this regard, UDCA stimulates the expression of the bile salt export pump proteins (BSEP), various drug resistance-associated proteins (MDR3 and MRP4), and promotes the elimination of the bile acids from hepatocytes.

[0196] In addition, another mechanism of protection of hepatocytes against the toxic bile acids is the inhibition of FA uptake by the transporters in the intestinal lumen due to the competition of UDCA for binding sites with the transporters.

[0197] Clinical studies on the effectiveness of UDCA have demonstrated a decrease in concentration level values of aminotransferases, alkaline phosphatase, and bilirubin in the patient blood serum, regardless of improvements in the processes at the histological level. Discussions remain regarding the effectiveness of UDCA in terms of overall patient survival, the impact on the need for liver transplantation, or the achievement of improvements at the histological level.

[0198] The debate about whether UDCA affects survival and mortality in the patients without liver transplantation is highlighted in review articles and meta-analyses, where scientists agree on some control of the biochemical markers in the patients taking UDCA, while the impact on histopathology and survival without transplantation cannot be assessed during study. However, the fact is that UDCA is not an ideal drug for the PBC treatment, and it does not always stop the disease progression.

[0199] In addition, asymptomatic cases of PBC have a favorable natural history and cannot be considered in the same cohort as symptomatic PBC. Of particular note, two out of three asymptomatic PBC patients benefit from the use of UDCA, compared with an equal or better history with reports of 57%, 70%, and over 90% of 10-year survival in the patients who received no treatment. Additionally, the expected median survival of the asymptomatic PBC patients was 10 and 16 years in two large cohorts observed for 24 years, while the median survival of the symptomatic patients was approximately 7 years.

[0200] A systematic review of 16 randomized clinical trials evaluating the efficacy of UDCA versus placebo in PBC showed no significant effect of UDCA on mortality or the need for liver transplantation in the patients. The effects of UDCA do not go beyond a certain improvement in the bilirubin level in the blood serum, liver tests of ALT and AST, and alkaline phosphatase. UDCA has not been found to improve the patient symptoms such as itching and fatigue, autoimmune conditions, liver histology, and portal pressure. In addition, the use of UDCA was associated to significant extend with the development of adverse events, such as weight gain.

[0201] Given these data, it can be concluded that, although UDCA has some therapeutic activity and proven efficacy, it is effective only in the early stages of PBC and in the asymptomatic patients.

[0202] Mechanism of action of obeticholic acid. OCA is a derivative of the primary human bile acid, chenodeoxycholic acid, and is the first agonist in its class to selectively bind to the nuclear farnesoid X receptor FXR. Due to chemical modification, the affinity of OCA to FXR is 100 times higher than that of chenodeoxycholic acid that is a natural FXR agonist. FXR is one of the key transcription factors that is overexpressed in the liver, kidneys, intestines, and adrenal glands and plays a key role in the pathogenesis of the inflammatory liver diseases. FXR has anti-inflammatory properties due to inhibition of the NF-κB gene. FXR regulates bile acid synthesis through two distinct mechanisms. In the liver, FXR increases the expression of the SHP (small heterodimer partner) gene. The SHP gene represses the transcription of the bile acid synthesis enzymes, such as CYP7A1 and CYP8B1, thus reducing the bile acid synthesis in hepatocytes. In addition, FXR regulates the activity of the bile acid transport systems in hepatocytes. In hepatocytes, FXR is involved in liver inflammation, fibrosis, regulation of metabolic pathways, and regeneration.

[0203] In addition to its central role in the bile acid metabolism, FXR activation also regulates the expression of various genes involved in glucose, lipid, and lipoprotein metabolism. Hepatic FXR inhibits fatty acid synthesis and absorption and enhances beta-oxidation by regulating lipid homeostasis.

[0204] OCA suppresses metabolic stress-induced activation of the p53 protein and apoptosis of damaged hepatocytes. Moreover, in addition to hepatocytes, OCA has been shown to exhibit anti-inflammatory and antifibrotic properties in immune cells, vascular smooth muscle cells, endothelial cells, and fibroblasts in vitro and in vivo.

[0205] It was also shown that stimulation of FXR in hepatic macrophage cells by OCA had an anti-inflammatory effect, inhibited the development of fibrosis, and stimulated liver tissue regeneration.

[0206] Comparison of the action of UDCA and OCA. As mentioned above, the main mechanisms of action of UDCA are immunomodulatory action, inhibition of absorption of hydrophobic FAs from the intestinal lumen and stimulation of FA release from hepatocytes, while for OCA these are modulation of FXR action, anti-inflammatory and antifibrotic effects, and inhibition of FA synthesis.

[0207] Based on the above scheme of 5 stages in the development of PBC (1—a trigger that triggers damage to liver cells; 2—development of cholestasis and induction of inflammation; 3—development of high-intensity inflammation; 4—fibrosis; 5—cirrhosis), today UDCA is used in the first two stages, since its anti-inflammatory properties are extremely weak, and OCA is used from the second to the last stage. Starting from the second stage of PBC, it is no longer rational to waste time treating the patient with UDCA pharmaceutical composition, since the percentage of the development and transition to irreversible stages of the disease is high. Therefore, an accurate and timely diagnosis of the PBC stage will lead to the possibility of prescribing safer and more effective treatment and use of a mixture/composition of two acids.

EMBODIMENTS OF THE INVENTION

[0208] To explore the possibility of using new markers for the PBC diagnosis, molecular and biological studies of the concentration level values of the markers of inflammation, apoptosis, and lipid peroxidation were carried out in the patients diagnosed with PBC and healthy volunteers. The results are listed below in Tables 1-3.

TABLE-US-00007 TABLE 1 Anthropomorphic data of healthy volunteers and patients diagnosed with PBC Healthy Patients volunteers with PBC (N = 58) (N = 60) Age (years) Mean value (years) 59.24 59.6 Range (years) 41-78 47-79 Sex Men (N) 10 5 Women (N) 48 55 Laboratory markers (mean value) ALT (U/l) 19.5 83 AST (U/l) 25.94 110 GGT (U/l) 23.4 210.5 Laboratory markers (mean value) AP (U/l) 87 354 Total bilirubin (μmol) 9.2 44.2 Presence of AMA antibodies (%) 0 98 IgM (g/l) 0.6 5.21

[0209] To determine the concentration level values of the markers in the blood serum, chemiluminescence and enzyme immunoassay methods were used.

TABLE-US-00008 TABLE 2 The concentration level values of inflammatory and apoptotic markers in the blood serum of healthy volunteers and patients diagnosed with PBC Healthy volunteers Patients with PBC TNF-α (ng/ml) 0.125 0.2975 IL-6 (pg/ml) 2.15 17.95 Nf-kB (pg/ml) 12.2 24.78 MCP-1/CCL2 (pg/ml) 212.2 375.9 Bcl-2 (U/ml) 0.22 0.33 MDA (nmol/ml) 4.21 16.98

[0210] Table 3 shows structured data about concentration level values of markers, which were identified on different stages of PBC.

TABLE-US-00009 TABLE 3 The concentration level values of inflammatory and apoptotic markers in the blood serum of healthy volunteers and patients by stages of PBC Healthy 1st 2nd 3rd 4th volunteers stage stage stage stage TNF-a (ng/ml) 0.125 0.18 0.25 0.34 0.42 IL-6 (pg/ml) 2.15 6.2 12.5 24.5 28.6 Nf-kB (pg/ml) 12.2 15.5 20.8 28.5 34.3 MCP-1/CCL2 212.2 250.5 311.2 457.3 484.6 (pg/ml) Bcl-2 (U/ml) 0.22 0.031 0.4 0.48 0.41 MDA (nmol/ml) 4.2 8.5 15.3 20.1 24.02

[0211] Based on the results obtained, it can be concluded that in the patients with PBC, there were found increased concentration level values of the markers of inflammation (TNF-α, IL-6, Nf-kB, and MCP-1), apoptosis (Bcl-2), as well as lipid peroxidation marker (MDA), in the blood.

[0212] Results Interpretation

[0213] TNF-α, IL-6, Nf-kB, MCP-1 are powerful markers of inflammation in tissues. Their increased activity indicates the acute phase of inflammation. In order to inhibit this inflammation, it is necessary to use strong anti-inflammatory drugs with local action. That is, the manifestation of such markers enables using not only UDCA, but switching immediately to the mixture of acids, such as UDCA and OCA.

[0214] Bcl-2 is mitochondrial apoptosis factor. The elevated concentration level values indicate pathological cell death. This pathological phenomenon can be eliminated by neutralizing the primary cause of the cell death (in this case, this is the toxic effect of the bile acids and the autoimmune process). Again, another marker that confirms the need to use the mixture: for the autoimmune process—UDCA, and to remove and stop the synthesis of fatty acids—UDCA and OCA together.

[0215] MDA is a marker of lipid peroxidation. Its increase indicates a high concentration of highly reactive oxygen in the tissues, which can be caused by both inflammation and the immune response.

[0216] In general, determination of the markers claimed by the authors enables achieving the following advantages:

[0217] 1) First of all, their use enables diagnosing PBC in very early stages, before the destruction of the liver cells and fibrosis begins, while biochemical markers (ALT/AST and bilirubin) show not the onset, but the progression of the disease. Successful diagnosis and early-stage treatment of the disease, in turn, enables bringing rates of adequate response to treatment in patients closer to 100%.

[0218] 2) Choosing the right treatment strategy. Without understanding the molecular pathogenesis, it is impossible to choose the correct individual treatment.

[0219] 3) More detailed and accurate tracking of the dynamics of the disease and more effective adjustments of the treatment based on the patient condition.

[0220] After obtaining a complete clinical picture of the disease based on the results of the study of the claimed markers, one can proceed directly to treatment.

[0221] To date, there is no pharmaceutical composition that is effective for the treatment of PBC in all stages of its course.

[0222] As a result of the studies on the use of the claimed markers in the PBC treatment, it was shown that in the patients diagnosed with PBC, there is an increase in the markers of inflammation and apoptosis, which indicates the feasibility of changing the protocols for the PBC treatment, in particular, the need for simultaneous use of UDCA and OCA in one dosage form.

[0223] For these purposes, a new all-purpose pharmaceutical composition for the PBC treatment, that comprises both ursodeoxycholic and obeticholic acids, and is effective for the use in all stages of PBC and has a complex mechanism of action, was developed. In particular, the use of the new pharmaceutical composition enables simultaneous blockage of the transport and synthesis of bile acids and achievement of a pronounced hepatoprotective effect.

[0224] According to some embodiments of the present disclosure, the claimed pharmaceutical composition may have the following composition:

TABLE-US-00010 Description Composition Embodiment Coated tablet Ursodeoxycholic acid - 100 mg 1 Obeticholic acid - 1 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 100 mg II Obeticholic acid - 2.5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 100 mg III Obeticholic acid - 5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 100 mg IV Obeticholic acid - 10 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 100 mg V Obeticholic acid - 20 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 100 mg VI Obeticholic acid - 50 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 250 mg VII Obeticholic acid - 1 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 250 mg VIII Obeticholic acid - 2.5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 250 mg IX Obeticholic acid - 5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 250 mg X Obeticholic acid - 10 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 250 mg XI Obeticholic acid - 20 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 250 mg XII Obeticholic acid - 50 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 500 mg XIII Obeticholic acid - 1 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 500 mg XIV Obeticholic acid - 2.5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 500 mg XV Obeticholic acid - 5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 500 mg XVI Obeticholic acid - 10 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 500 mg XVII Obeticholic acid - 20 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 500 mg XVIII Obeticholic acid - 50 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 1000 mg XIX Obeticholic acid - 1 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 1000 mg XX Obeticholic acid - 2.5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 1000 mg XXI Obeticholic acid - 5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 1000 mg XXII Obeticholic acid - 10 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 1000 mg XXIII Obeticholic acid - 20 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 1000 mg XXIV Obeticholic acid - 50 mg Excipient

[0225] According to preferred embodiments of the present disclosure, the claimed pharmaceutical composition may have the following composition:

TABLE-US-00011 Embodiment Coated tablet Ursodeoxycholic acid - 150 mg XXX Obeticholic acid - 1 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 250 mg XXXI Obeticholic acid - 1 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 250 mg XXXII Obeticholic acid - 1.25 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 150 mg XXXIII Obeticholic acid - 2 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 250 mg XXXIV Obeticholic acid - 2.5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 500 mg XXXV Obeticholic acid - 2.5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 750 mg XXXVI Obeticholic acid - 2.5 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 150 mg XXXVII Obeticholic acid - 5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 250 mg XXXVIII Obeticholic acid - 5 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 500 mg XXXIX Obeticholic acid - 5 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 750 mg XXXVII Obeticholic acid - 5 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 150 mg XL Obeticholic acid - 10 mg Excipient Embodiment Coated tablet Ursodeoxycholic acid - 250 mg XLI Obeticholic acid - 10 mg Excipient Embodiment Uncoated tablet Ursodeoxycholic acid - 500 mg XLI Obeticholic acid - 10 mg Excipient Embodiment Capsule Ursodeoxycholic acid - 750 mg XLII Obeticholic acid - 10 mg Excipient

[0226] Above-mentioned embodiments can be produced, for example, in the following ways.

[0227] Method 1.

[0228] Step 1: Weighing. Weigh the required amount of the active and auxiliary components.

[0229] Step 2: Sifting of the raw materials. Sift the raw materials through a sieve with a specific mesh size, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0230] Step 3: Dry mixing. Transfer the sifted materials to a dry mixing apparatus and mix for a specific amount of time at a specific mixing speed in accordance with the technical instructions for the manufacture of the solid dosage form to produce mixed powder.

[0231] Step 4: Producing a slurry. Mix a specific volume of the purified water EP having a specific temperature with a base to produce a slurry.

[0232] Step 5: Blending and bonding. Granulation. Mix the slurry produced in Step 4 with the powder from Step 3 in the granulator.

[0233] Step 6: Semi-drying. Transfer the granules produced in Step 5 to the drying apparatus and dry at a specific temperature for a specific amount of time, in accordance with the technical instructions for the manufacture of the solid dosage form to produce dried granules.

[0234] Step 7: Crushing and sifting all dried granules from Step 6 to produce sifted granules.

[0235] Step 8: Final drying. Transfer the sifted granules produced in Step 7 to the drying apparatus and dry at a specific temperature for a specific amount of time, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0236] Step 9: Lubricating.

[0237] Transfer all granules produced in Step 8 to a blender and mix for a specific amount of time. Add a material to lubricate the granules and mix for a specific amount of time.

[0238] Step 10: Tablet compression.

[0239] Step 11: Tablet coating.

[0240] Method 2.

[0241] Step 1: Weighing. Weigh the required amount of the active and auxiliary components.

[0242] Step 2: Sifting of the raw materials. Sift the raw materials through a sieve with a specific mesh size, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0243] Step 3: Dry mixing. Transfer the sifted materials to a dry mixing apparatus and mix for a specific amount of time at a specific mixing speed in accordance with the technical instructions for the manufacture of the solid dosage form to produce mixed powder.

[0244] Step 4: Adding granulating solution and wetting.

[0245] Add pre-prepared granulating solution to the mixed powder produced in Step 3 and mix to produce mixture.

[0246] Step 5: Transfer the mixture produced in Step 4 to the granulator and mix to produce granules.

[0247] Step 6: Drying. Transfer the granules produced in Step 5 to the drying apparatus and dry at a specific temperature for a specific amount of time, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0248] Step 7: Granule coating.

[0249] Method 3.

[0250] Step 1: Weighing. Weigh the required amount of the active and auxiliary components.

[0251] Step 2: Sifting of the raw materials. Sift the raw materials through a sieve with a specific mesh size, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0252] Step 3: Dry mixing. Transfer the sifted materials to a dry mixing apparatus and mix for a specific amount of time at a specific mixing speed in accordance with the technical instructions for the manufacture of the solid dosage form to produce mixed powder.

[0253] Step 4: Preparing of the granulating solution. Add the granulating solution base and purified water EP into the preparation container and mix for a specific amount of time in accordance with the technical instructions for the manufacture of the solid dosage form to produce granulating solution.

[0254] Step 5: Adding of the granulating solution and wetting.

[0255] To the dry mixture produced in Step 3, add the granulating solution produced in Step 4 and mix to produce mixture.

[0256] Step 6: Transfer the mixture produced in Step 5 to the granulator and mix to produce granules.

[0257] Step 7: Drying. Transfer the granules produced in Step 6 to the drying apparatus and dry at a specific temperature for a specific amount of time, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0258] Step 8: Granule coating.

[0259] Method 4.

[0260] Step 1: Weighing. Weigh the required amount of the active and auxiliary components.

[0261] Step 2: Sifting of the raw materials. Sift the raw materials through a sieve with a specific mesh size, in accordance with the technical instructions for the manufacture of the solid dosage form.

[0262] Step 3: Dry mixing. Transfer the sifted materials to a dry mixing apparatus and mix for a specific amount of time at a specific mixing speed in accordance with the technical instructions for the manufacture of the solid dosage form to produce mixed powder.

[0263] Step 4: Tablet compression.

[0264] Step 5: Tablet coating.

[0265] As an excipient claimed pharmaceutical can comprise any excipients acceptable for use in pharmaceutical industry, in particular, excipients that are used to produce solid dosage forms, such as fillers, binders, antiadherents, glidants, lubricants, disintegrants, plasticizer, sweeteners, opaquants, flavorants.

[0266] As used herein, the term “filler” is intended to mean inert substances used to create the desired bulk, flow properties, and compression characteristics in the preparation of tablets and capsules. Exemplary fillers include, but not limited to, dibasic calcium phosphate, kaolin, lactose, sucrose, mannitol, cellulose and its derivatives, precipitated calcium carbonate, sorbitol, and starch.

[0267] As used herein, the term “binder” is intended to mean a substance used to cause adhesion of powder particles in granulations. Exemplary binders include, but not limited to, acacia, tragacanth, alginic acid and salts thereof, gelatin, cellulose and its derivatives, poly(vinylpyrrolidone), liquid glucose, povidone, polyethylene glycol, polypropylene glycol, polyoxyethylene-polypropylene copolymer, polyethylene ester, polyethylene sorbitan ester, polyethylene oxide, guar gum, polysaccharide, bentonites, sugars, invert sugars, compressible sugar, poloxamers, collagen, albumin, and starch and its derivatives.

[0268] As used herein, the term “antiadherent” is intended to mean an agent that prevents the sticking of tablet formulation ingredients to punches and dies in a tableting machine during production. Exemplary antiadherents include, but not limited to, magnesium stearate, talc, calcium stearate, glyceryl behenate, polyethylene glycol (PEG), hydrogenated vegetable oil, mineral oil, stearic acid.

[0269] As used herein, the term “glidant” is intended to mean an agent used in tablet and capsule formulations to promote flowability of the granulation. Exemplary glidants include, but not limited to, colloidal silica, cornstarch, talc, calcium silicate, magnesium silicate, colloidal silicon, silicon hydrogel.

[0270] As used herein, the term “lubricant” is intended to mean a substance used to reduce friction during compression or other processing. Exemplary lubricants include, but not limited to, calcium stearate, magnesium stearate, mineral oil, stearic acid, and zinc stearate.

[0271] As used herein, the term “disintegrant” is intended to mean a compound used to promote the disruption of the solid mass into smaller particles that are more readily dispersed or dissolved. Exemplary disintegrants include, but not limited to, starches such as corn starch, potato starch, pre-gelatinized and modified starches thereof, clays, such as bentonite, cellulose and its derivatives, such as microcrystalline cellulose, carboxymethylcellulose calcium, cellulose polyacrilin potassium, alginates, sodium starch glycolate, gums such as agar, guar, locust bean, karaya, pectin, tragacanth; crospovidone.

[0272] As used herein, the term “plasticizer” includes compounds capable of plasticizing or softening a polymer or binder used in invention by way of lowering the melting temperature or glass transition temperature of the polymer or binder. Exemplary plasticizers include, but not limited to, low molecular weight polymers, oligomers, copolymers, oils, low molecular weight polyols, multi-block polymers, single block polymers, and glycerin.

[0273] As used herein, the term “sweetener” is intended to mean a compound used to impart sweetness. Exemplary sweeteners include, but not limited to, aspartame, dextrose, glycerin, mannitol, saccharin sodium, sorbitol and sucrose.

[0274] As used herein, the term “opaquant” is intended to mean a compound used to render a tablet coating opaque. Exemplary opaquant include, but not limited to, titanium dioxide, talc.

[0275] As used herein, the term “flavorant” is intended to mean a compound used to impart a pleasant flavor and often odor. Exemplary flavorants include, but not limited to, synthetic flavor oils and flavoring aromatics and/or natural oils, extracts from plants, leaves, flowers, fruits and combinations thereof, and other substances known to one skilled in the art.

[0276] Method A.

[0277] Step 1. Weighing. Weigh the required amount of the active and auxiliary components (per 100 kg of the total mixture).

[0278] 1) UDCA+OCA—80 kg.

[0279] 2) Corn starch—15 kg.

[0280] 3) Lactose—3 kg.

[0281] 4) Povidone—2 kg.

[0282] Step 2. Sifting of the raw materials. Sift the raw materials through a sieve with a mesh size of at least 30 mesh.

[0283] Step 3. Dry mixing. Transfer the sieved materials to a dry mixer and mix them for 1 hour.

[0284] Step 4. Transfer the mixture produced in Step 3 to a compression machine, and begin the process of producing the tablets by direct compression.

[0285] Step 5. Transfer the tablets produced in Step 4 to a tablet film coating machine. Coating of the tablets with the film achieved by immersion in an ethanol solution of a film former (eudragit E).

[0286] Step 6. Drying of the tablets produced.

[0287] Method B.

[0288] Step 1. Weighing. Weigh the required amount of the active and auxiliary components (per 150 kg of the total mixture).

[0289] 1) UDCA+OCA—112.5 kg.

[0290] 2) Corn starch—16.5 kg.

[0291] 3) Lactose—18.0 kg.

[0292] 4) Povidone—3.0 kg.

[0293] Step 2. Sifting of the raw materials. Sift the raw materials through a sieve with a mesh size of at least 30 mesh.

[0294] Step 3. Dry mixing. Transfer the sieved materials to a dry mixer and mix them for 1 hour.

[0295] Step 4: Transfer the mixture produced in Step 3 to a compression machine, and begin the process of producing the tablets by direct compression.

[0296] Step 5. Transfer the tablets produced in Step 4 to a tablet film coating machine. Coating of the tablets with the film achieved by immersion in an ethanol solution of a film former (eudragit L).

[0297] Step 6. Drying of the tablets produced.

[0298] Method C Step 1. Weighing. Weigh the required amount of the active and auxiliary components (per 200 kg of the total mixture).

[0299] 1) UDCA+OCA—150 kg.

[0300] 2) Microcrystalline cellulose—30 kg.

[0301] 3) Lactose—18 kg.

[0302] 4) Povidone—2 kg.

[0303] Step 2. Sifting of the raw materials. Sift the raw materials through a sieve with a mesh size of at least 30 mesh.

[0304] Step 3. Dry mixing. Transfer the sifted materials to a dry mixer and mix them for 1 hour.

[0305] Step 4: Transfer the mixture produced in Step 3 to a compression machine, and begin the process of producing the tablets by direct compression.

[0306] Step 5. Transfer the tablets produced in Step 4 to a tablet film coating machine. Coating of the tablets with the film achieved by immersion in an ethanol solution of a film former (eudragit L).

[0307] Step 6. Drying of the tablets produced.

[0308] Clinical Studies

[0309] A double-blind clinical study was conducted on the effectiveness of the combined pharmaceutical composition comprising UDCA and OCA compared with the monocomposition comprising only UDCA for the patients with the following concentration level values of the claimed group of the markers (corresponding to 1 Stage of PBC): [0310] IL-6—at least 10%; [0311] Nf-kB—at least 5%; [0312] MCP-1/CCL2—at least 5%; [0313] Bcl-2—at least 5%; [0314] TNF-α—at least 12%; [0315] MDA—at least 0.5%.

TABLE-US-00012 TABLE 4 Data on the tested groups before the treatment Group Group “UDCA Group “UDCA “UDCA 15 15 mg/kg + 15 mg/kg + mg/kg” OCA 5 mg” OCA 10 mg” (N = 68) (N = 65) (N = 63) Age (years) Mean value (years) 59.6 58.2 59.1 Range (years) 47-79 47-78 48-77 Sex Men (N) 9 7 3 Women (N) 59 58 60 Laboratory markers (mean concentration level values) ALT (U/l) 82.7 71.33 84.3 AST (U/l) 105.2 106.9 112.5 GGT (U/l) 207.5 212.3 208.8 AP (U/l) 368 349 325 Total bilirubin 40.3 38.6 41.33 (μmol/l) Presence of AMA 95 98 94 antibodies (%) IgM (g/l) 0.5 0.7 0.58 TNF-α (ng/ml) 0.33 0.35 0.33 IL-6 (pg/ml) 23.54 24.87 26.2 Nf-kB (pg/ml) 65.5 67.6 69.5 MCP-1/CCL2 (pg/ml) 468.5 485.4 495.2 Bcl-2 (U/ml) 0.72 0.69 0.71 MDA (nmol/ml) 23.08 22.58 24.01

[0316] Comparison of the effectiveness of the combined (UDCA+OCA) pharmaceutical composition and monocomposition (UDCA) was carried out for 6 and 12 months. The patients diagnosed with PBC were divided into 3 groups depending on the treatment regimen:

[0317] 1) Group UDCA 15 mg/kg;

[0318] 2) Group UDCA 15 mg/kg+OCA 5 mg;

[0319] 3) Group UDCA 15 mg/kg+OCA 10 mg.

[0320] After the treatment, venous blood samples were taken from the patients and the concentrations of biochemical and molecular markers were determined.

[0321] As markers of the treatment effectiveness, two groups of markers were used: biochemical and molecular biological.

[0322] The first group included: ALT, AST, GGT (Gamma-glutamyl Transferase), AP and total bilirubin.

[0323] The second group included: TNF-α, IL-6, Nf-kB, MCP-1/CCL2, BCL-2, and MDA.

[0324] The study results are shown in Tables 5-6.

TABLE-US-00013 TABLE 5 Concentration level values of the markers in the blood plasma (6 months) UDCA UDCA UDCA 15 mg/kg + 15 mg/kg + 15 mg/kg OCA 5 mg OCA 10 mg (N = 68) (N = 65) (N = 63) Before After Before After Before After ALT (U/l) 55.7 37.5 56.33 28.2 58.3 25.3 AST (U/l) 58.6 39.4 59.5 31.7 61.4 25.1 GGT (U/l) 207.5 135.2 212.3 74.2 208.8 60.2 AP (U/l) 297.2 190 284.5 147.2 281.2 120.2 Total 57.6 20 59.8 12.8 54.3 12.1 bilirubin (μmol/l) TNF-α 0.33 0.29 0.35 0.123 0.33 0.1 (ng/ml) IL-6 (pg/ml) 23.54 20.2 24.87 5.15 26.2 3.15 Nf-kB (pg/ml) 65.5 60.2 67.6 15.2 69.5 12.3 MCP-1/ 468.5 412.3 485.4 212.8 495.2 200.2 CCL2 (pg/ml) Bcl-2 (U/ml) 0.42 0.4 0.39 0.3 0.41 0.28 MDA 23.08 22.01 22.58 10.17 24.01 7.2 (nmol/ml)

TABLE-US-00014 TABLE 6 Changes of the marker concentration in the blood plasma (6 months), in % of the initial value UDCA UDCA UDCA 15 mg/kg + 15 mg/kg + 15 mg/kg OCA 5 mg OCA 10 mg (N = 68) (N = 65) (N = 63) Before After Before After Before After ALT 100 67.32 100 50.06 100 43.40 AST 100 67.24 100 53.28 100 40.88 GGT 100 65.16 100 34.95 100 28.83 AP 100 63.93 100 51.74 100 42.75 Total 100 34.72 100 21.40 100 22.28 bilirubin TNF-α 100 87.88 100 35.14 100 30.30 IL-6 100 85.81 100 20.71 100 12.02 Nf-kB 100 91.91 100 22.49 100 17.70 MCP-1/ 100 88.00 100 43.84 100 40.43 CCL2 Bcl-2 100 95.24 100 76.92 100 68.29 MDA 100 95.36 100 45.04 100 29.99

[0325] Results of the study on the effectiveness of the treatment of the patients with UDCA or UDCA+OCA (12 months) are shown in the tables below.

TABLE-US-00015 TABLE 7 Concentration level values of the markers in the blood plasma (12 months) UDCA UDCA UDCA 15 mg/kg + 15 mg/kg + 15 mg/kg OCA 5 mg OCA 10 mg (N = 68) (N = 65) (N = 63) Before After Before After Before After ALT (U/l) 82.7 55.67 71.33 35.7 84.3 36.59 AST (U/l) 105.2 70.73 106.9 56.96 112.5 45.99 GGT (U/l) 207.5 135.2 212.3 74.2 208.8 60.2 AP (U/l) 368.4 235.36 349 180.57 325 138.93 Total 40.3 26.03 38.6 14.74 41.33 9.2 bilirubin (μmol/l) TNF-α 0.33 0.29 0.35 0.123 0.33 0.1 (ng/ml) IL-6 (pg/ml) 23.54 20.2 24.87 5.15 26.2 3.15 Nf-kB (pg/ml) 65.5 60.2 67.6 15.2 69.5 12.3 MCP-1/ 468.5 412.3 485.4 212.8 495.2 200.2 CCL2 (pg/ml) Bcl-2 (U/ml) 0.72 0.52 0.69 0.43 0.71 0.34 MDA 23.08 22.01 22.58 10.17 24.01 7.2 (nmol/ml)

TABLE-US-00016 TABLE 8 Changes of the marker concentration in the blood plasma (12 months), in % of the initial value UDCA UDCA UDCA 15 mg/kg + 15 mg/kg + 15 mg/kg OCA 5 mg OCA 10 mg (N = 68) (N = 65) (N = 63) Before After Before After Before After ALT 100 67.32 100 50.06 100 43.40 AST 100 67.24 100 53.28 100 40.88 GGT 100 65.16 100 34.95 100 28.83 AP 100 63.93 100 51.74 100 42.75 Total 100 64.58 100 38.2 100 22.28 bilirubin TNF-α 100 87.88 100 35.14 100 30.30 IL-6 100 85.81 100 20.71 100 12.02 Nf-kB 100 91.91 100 22.49 100 17.70 MCP-1/ 100 88.00 100 43.84 100 40.43 CCL2 Bcl-2 100 72 100 62.32 100 47.89 MDA 100 95.36 100 45.04 100 29.99

[0326] The results obtained enable drawing the following conclusions:

[0327] 1. Ursodeoxycholic acid monocomposition was more effective in reducing biochemical but not molecular and biological markers of inflammation and apoptosis.

[0328] 2. The effect of ursodeoxycholic acid on the markers of inflammation, apoptosis and lipid peroxidation was insignificant.

[0329] 3. The use of the combined preparations enables reducing not only biochemical markers, but also coping effectively with an acute inflammatory process and ceasing the cell death and development of fibrosis.

[0330] More detailed study results per months are given below in Tables 9-14.

TABLE-US-00017 TABLE 9 Concentration level values of the markers in the blood plasma when using the standard treatment regimen No. 1 (UDCA 15 mg/kg) Months 0 1 3 6 9 12 ALT (U/l) 82.7 80.4 74.81 68.2 62.47 55.67 AST (U/l) 105.2 100.7 87.3 81.4 74.5 70.73 GGT (U/l) 207.5 198 178.8 156.2 141.5 135.2 AP (U/l) 368.4 352.5 290.6 269.45 247.5 235.36 Total 40.3 39.8 33.8 32.8 28.03 26.03 bilirubin (μmol/l) TNF-α 0.33 0.33 0.324 0.315 0.3 0.29 (ng/ml) IL-6 (pg/ml) 23.54 23.35 22.95 22 21.48 20.2 Nf-kB (pg/ml) 65.5 64.4 64.1 62.8 61.4 60.2 MCP-1/ 468.5 465.4 448 431.4 419.7 412.3 CCL2 (pg/ml) Bcl-2 (U/ml) 0.72 0.68 0.62 0.58 0.54 0.52 MDA 23.08 22.87 22.54 22.37 22.12 22.01 (nmol/ml)

TABLE-US-00018 TABLE 10 Changes of the marker concentration in the blood plasma when using the standard treatment regimen No. 1 (UDCA 15 mg/kg), in % of the initial value Months 0 1 3 6 9 12 ALT 100.00 97.22 90.46 82.47 75.54 67.32 AST 100.00 95.72 82.98 77.38 70.82 67.23 GGT 100.00 95.42 86.17 75.28 68.19 65.16 AP 100.00 97.20 88.10 74.45 69.18 63.93 Total bilirubin 100.00 96.18 87.50 77.60 70.49 64.58 TNF-α 100.00 100.00 98.18 95.45 90.91 87.88 IL-6 100.00 99.19 97.49 93.46 91.25 85.81 Nf-kB 100.00 98.32 97.86 95.88 93.74 91.91 MCP-1/CCL2 100.00 99.34 95.62 92.08 89.58 88.00 Bcl-2 100.00 94.44 86.11 80.56 75.00 72.22 MDA 100.00 99.09 97.66 96.92 95.84 95.36

[0331] The above data show the change in the concentration level values of biochemical and molecular markers of the PBC development, when using treatment regimen No. 1. It can be seen that the chosen treatment regimen was the most effective in reducing markers, such as ALT, AST, GGT, AP, total bilirubin and BCL-2, which indicates that such a treatment regimen is somewhat effective, but it has little effect on inflammation processes.

TABLE-US-00019 TABLE 11 Concentration level values of the markers in the blood plasma when using the provided treatment regimen No. 2 (UDCA 15 mg/kg + OCA 5 mg). Months 0 1 3 6 9 12 ALT (U/l) 71.33 69.7 61.7 52.75 41.79 35.7 AST (U/l) 106.9 101.4 87 79.45 71.7 56.96 GGT (U/l) 212.3 207.4 184.4 134.6 87.5 74.2 AP (U/l) 349 332.3 285.6 245.8 210.5 180.57 Total 38.6 37.6 31.05 24.8 16.87 14.74 bilirubin (μmol/l) TNF-α 0.35 0.27 0.24 0.178 0.14 0.123 (ng/ml) IL-6 (pg/ml) 24.87 18.6 16.7 12.6 8.9 5.15 Nf-kB (pg/ml) 67.6 59.7 41.5 29.7 19.4 15.2 MCP-1/ 485.4 403.6 301.5 270 235.4 212.8 CCL2 (pg/ml) Bcl-2 (U/ml) 0.69 0.67 0.62 0.51 0.45 0.43 MDA 22.58 19.68 17.8 13.8 12.6 10.17 (nmol/ml)

TABLE-US-00020 TABLE 12 Changes of the marker concentration in the blood plasma when using the provided treatment regimen No. 2 (UDCA 15 mg/kg + OCA 5 mg), in % of the initial value Months 0 1 3 6 9 12 ALT 100.00 97.71 86.50 73.95 58.59 50.05 AST 100.00 94.86 81.38 74.32 67.07 53.28 GGT 100.00 97.69 86.86 63.40 41.22 34.95 AP 100.00 95.60 84.91 70.29 57.32 51.74 Total bilirubin 100.00 98.16 76.76 59.87 49.16 38.19 TNF-α 100.00 77.14 68.57 50.86 40.00 35.14 IL-6 100.00 74.79 67.15 50.66 35.79 20.71 Nf-kB 100.00 88.31 61.39 43.93 28.70 22.49 MCP-1/CCL2 100.00 83.15 62.11 55.62 48.50 43.84 Bcl-2 100.00 97.10 89.86 73.91 65.22 62.32 MDA 100.00 87.16 78.83 61.12 55.80 45.04

[0332] The above data show the change in the concentration level values of biochemical and molecular markers of the PBC development, when using treatment regimen No. 2. It can be seen that the selected treatment regimen was more effective than regimen No. 1.

[0333] In addition to the fact that in this case there was a more significant decrease in biochemical markers characterizing the general functional condition of the liver, there was also a significant decrease in the markers of inflammation, chemoattraction of monocytes from peripheral blood, apoptosis and lipid peroxidation.

TABLE-US-00021 TABLE 13 Concentration level values of the markers in the blood plasma when using the provided treatment regimen No. 3 (UDCA 15 mg/kg + OCA 10 mg). Months 0 1 3 6 9 12 ALT (U/l) 84.3 82.4 71.9 59.1 41.8 36.59 AST (U/l) 112.5 109.6 85.6 69.96 51.8 45.99 GGT (U/l) 208.8 198.6 135 106 89.5 60.2 AP (U/l) 325 319.5 289.57 210.5 178.57 138.93 Total 41.33 39.25 31.05 24.6 16.8 9.2 bilirubin (μmol/l) TNF-α 0.33 0.28 0.25 0.18 0.13 0.1 (ng/ml) IL-6 (pg/ml) 26.2 21.3 16.9 13.69 7.45 3.15 Nf-kB (pg/ml) 69.5 57.61 35.6 22 14.6 12.3 MCP-1/CCL2 495.2 401.7 301.8 270.2 230.8 200.2 (pg/ml) Bcl-2 (U/ml) 0.71 0.68 0.64 0.52 0.46 0.34 MDA 24.01 21.5 18.6 13.8 7.92 7.2 (nmol/ml)

TABLE-US-00022 TABLE 14 Changes of the marker concentration in the blood plasma when using the provided treatment regimen No. 3 (UDCA 15 mg/kg + OCA 10 mg), in % of the initial value Months 0 1 3 6 9 12 ALT 100.00 97.75 85.29 70.11 49.58 43.40 AST 100.00 97.42 76.09 62.19 46.04 40.88 GGT 100.00 95.11 64.66 50.77 42.86 28.83 AP 100.00 98.67 74.32 62.19 47.21 42.75 Total bilirubin 100.00 96.50 65.56 36.10 29.10 22.28 TNF-α 100.00 84.85 75.76 54.55 39.39 30.30 IL-6 100.00 81.30 64.50 52.25 28.44 12.02 Nf-kB 100.00 82.89 51.22 31.65 21.01 17.70 MCP-1/CCL2 100.00 81.12 60.95 54.56 46.61 40.43 Bcl-2 100.00 95.77 90.14 73.24 64.79 47.89 MDA 100.00 89.55 77.47 57.48 32.99 29.99

[0334] The above data show the change in the concentration level values of biochemical and molecular markers of the PBC development, when using treatment regimen No. 3. FIG. 3 shows that the selected treatment regimen was more effective than regimens No. 1 and No. 2.

[0335] When using this regimen, there was a significant decrease in biochemical markers characterizing the general functional condition of the liver, as well as a significant decrease in the markers of inflammation, chemoattraction of monocytes from peripheral blood, apoptosis and lipid peroxidation.

[0336] If we compare all three treatment regimens, we can say that:

[0337] 1) UDCA monocomposition has a specific effect on the overall functional condition of the liver, but almost no effect on the inflammatory and immunological component of the disease.

[0338] 2) The combined pharmaceutical composition containing UDCA and OCA is more effective in the treatment of PBC, because it affects not only the general functional condition of the liver, but also the molecular mechanisms of the development of this disease, that is, the treatment is not only symptomatic.

[0339] 3) The use of the new markers enables not only developing the new treatment regimens with the combined pharmaceutical composition, but also evaluating the effectiveness of the treatment in more detail.

[0340] Solid Dosage Forms

[0341] Specifically, the authors of the present disclosure suggest the manufacture of the claimed pharmaceutical composition comprising obeticholic and ursodeoxycholic acids in the solid dosage form, for example, in the form of a tablet or capsule.

[0342] The development of the new all-purpose solid dosage form for the treatment of PBC, that comprises both ursodeoxycholic and obeticholic acids and is characterized by a complex mechanism of action, enables achieving effective treatment of all stages of PBC. In particular, the use of the new solid dosage form enables simultaneous blockage of the transport and synthesis of bile acids, and achievement of a pronounced hepatoprotective effect.

[0343] Based on the studies conducted, the authors consider it necessary to use the combined solid dosage form for the treatment of PBC, the active components of which are ursodeoxycholic acid, that stimulates the excretion of bile acids, and obeticholic acid, that stimulates the nuclear FXR receptor, which leads to inhibition of bile acid synthesis, anti-inflammatory, antifibrotic and antiapoptotic effect.

[0344] A study of the pharmacokinetics of obeticholic acid in a monocom position and as part of a composition with ursodeoxycholic acid when taken orally was conducted. It was discovered that when taking OCA with UDCA, UDCA inhibits the absorption of OCA, reducing bioavailability from 89% to 56% for 5 mg OCA, and from 91% to 61% for 10 mg OCA, respectively. The results of the pharmacokinetic studies are shown in Table 15.

TABLE-US-00023 TABLE 15 Pharmacokinetic parameters of the compositions with OBA and UDCA, compared with the solid dosage form according to the present disclosure OCA 5 mg + OCA 10 mg + OCA 5 mg + OCA 10 mg + UDCA 500 mg UDCA 500 mg OCA OCA UDCA UDCA enteric coating, enteric coating, 5 mg 10 mg 500 mg 500 mg modified release modified release Cmax (ng/ml) 16.7 26.4 7.1 14.8 16.2 26.8 AUCt (ng*h/ml) 34.8 66.1 19.4 35.8 33.7 65.8 Tmax (min) 40 90 95 140 45 87 Bioavailability 89 91 56 61 87 88 (%)

[0345] Thus, to improve the therapeutic parameters of the combined solid dosage form with ursodeoxycholic and obeticholic acids, the authors of the present disclosure developed the dosage form of the multilayer modified release tablet. This once again leads to increased compliance of the claimed pharmaceutical composition and solid dosage form and provides additional arguments in favor of the pharmaceutical composition with OCA and UDCA in one dosage form of the tablet with a structural solution. Providing the complexity of the treatment (many different pills) and the specifics of the symptoms with manifestations of fatigue and lack of concentration, the patients, when taking two separate pharmaceutical compositions, will make mistakes in taking medications: both in the number of the tablets and in taking them in time. Moreover, the problem of competition between OCA and UDCA for the transporters was investigated only now by the authors of the present disclosure and was not previously considered in this context. And if the patient takes several different pharmaceutical compositions with a discrete separate content of UDCA and OCA, this will lead to a decrease in the effectiveness of the treatment due to the suppression of OCA by ursodeoxycholic acid.

[0346] One of the solutions suggested by the authors is the multilayer tablet, in which the release of OCA occurs first, and only after that the release of UDCA occurs.

[0347] According to one of the embodiments of the present disclosure, the tablet consists of the following layers:

[0348] 1) The outer layer that can be formulated to be enteric (resistant to hydrochloric acid due to special polymers).

[0349] 2) The second layer comprising obeticholic acid (OCA Layer). According to one of the embodiments, after the dissolution of the outer layer, the release and absorption of OCA occurs in the intestinal lumen.

[0350] 3) The STOP layer. It consists of a special layer, the swelling and dissolution of which takes about 1-2 hours, which allows the OCA to be completely absorbed. After the dissolution of this layer, the release of UDCA begins.

[0351] 4) The core comprising ursodeoxycholic acid. After the dissolution of all the above layers, UDCA is released and absorbed. Potentially, this layer could be designed with extended release with a release time of up to 6 hours.

[0352] According to another embodiment of the present disclosure, the tablet consists of the following layers:

[0353] 1) The outer layer, that may be immediate release coating.

[0354] 2) The second layer comprising obeticholic acid (OCA Layer).

[0355] 3) The layer that is enteric coating.

[0356] 4) The core comprising ursodeoxycholic acid, that can be designed with extended release with UDCA release time up to 6 hours.

[0357] Some potential embodiments of the claimed solid dosage forms in the tablet form are presented below in Table 16.

TABLE-US-00024 TABLE 16 Embodiments of the claimed solid dosage forms in the tablet form Description Composition Embodiment 1 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release STOP-layer Layer No. 3 - obeticholic acid - 1 mg Outer layer - enteric coating Embodiment 2 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 3 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release STOP-layer Layer No. 3 - obeticholic acid - 5 mg Outer layer - enteric coating Embodiment 4 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 5 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release STOP-layer Layer No. 3 - obeticholic acid - 20 mg Outer layer - enteric coating Embodiment 6 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating Embodiment 7 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release STOP-layer Layer No. 3 - obeticholic acid - 1 mg Outer layer - enteric coating Embodiment 8 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 9 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release STOP-layer Layer No. 3 - obeticholic acid - 5 mg Outer layer - enteric coating Embodiment 10 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 11 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release STOP-layer Layer No. 3 - obeticholic acid - 20 mg Outer layer - enteric coating Embodiment 12 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating Embodiment 13 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release STOP-layer Layer No. 3 - obeticholic acid - 1 mg Outer layer - enteric coating Embodiment 14 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 15 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release STOP-layer Layer No. 3 - obeticholic acid - 5 mg Outer layer - enteric coating Embodiment 16 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 17 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release STOP-layer Layer No. 3 - obeticholic acid - 20 mg Outer layer - enteric coating Embodiment 18 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 19 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release STOP-layer Layer No. 3 - obeticholic acid - 1 mg Outer layer - enteric coating Embodiment 20 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 21 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release STOP-layer Layer No. 3 - obeticholic acid - 5 mg Outer layer - enteric coating Embodiment 22 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 23 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000mg release STOP-layer Layer No. 3 - obeticholic acid - 20 mg Outer layer - enteric coating Embodiment 24 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating Embodiment 25 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release Enteric coating Layer No. 3 - obeticholic acid - 1 mg Outer layer - film non-enteric coating Embodiment 26 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 27 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release Enteric coating Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 28 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 29 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release Enteric coating Layer No. 3 - obeticholic acid - 20 mg Outer layer - film non-enteric coating Embodiment 30 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 100 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating Embodiment 31 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release Enteric coating Layer No. 3 - obeticholic acid - 1 mg Outer layer - film non-enteric coating Embodiment 32 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 33 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release Enteric coating Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 34 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 35 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release Enteric coating Layer No. 3 - obeticholic acid - 20 mg Outer layer - film non-enteric coating Embodiment 36 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 250 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating Embodiment 37 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release Enteric coating Layer No. 3 - obeticholic acid - 1 mg Outer layer - film non-enteric coating Embodiment 38 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 39 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release Enteric coating Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 40 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 41 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release Enteric coating Layer No. 3 - obeticholic acid - 20 mg Outer layer - film non-enteric coating Embodiment 42 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 43 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release Enteric coating Layer No. 3 - obeticholic acid - 1 mg Outer layer - film non-enteric coating Embodiment 44 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 2.5 mg Outer layer - film non-enteric coating Embodiment 45 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release Enteric coating Layer No. 3 - obeticholic acid - 5 mg Outer layer - film non-enteric coating Embodiment 46 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating Embodiment 47 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1002 mg release Enteric coating Layer No. 3 - obeticholic acid - 20 mg Outer layer - film non-enteric coating Embodiment 48 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 1000 mg release and extended Enteric coating release of UDCA Layer No. 3 - obeticholic acid - 50 mg Outer layer - film non-enteric coating

[0358] According to some embodiments of the present disclosure, the claimed solid dosage form may have the following composition:

Example 50

[0359]

TABLE-US-00025 Unit Composition UDCA + OCA combination Strength 500 + 25 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 500 Disodium Phosphate 50 Polyvinylpyrrolidone 24 Microcrystalline Cellulose 150 Magnesium Stearate 10 Talc 6 Hydroxypropylmethylcellulose 14 PEG 6000 10 Titanium Dioxide 2 Cellulose Acetate Phthalate 24 Acetylated Monoglycerides 4 STOP Layer Microcrystalline Cellulose 12 Starch maize 6 Crospovidone 4 Polyvinylpyrrolidone 4 OCA Layer Obeticholic acid 25 Microcrystalline Cellulose 29 Sodium Starch Glycollate 8 Colloidal Silicon Dioxide 2 Magnesium Stearate 4 Gastric acid soluble coating Opadry (Ready Film coating material) 12

Example 51

[0360]

TABLE-US-00026 Unit Composition UDCA + OCA combination Strength 100 + 5 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 100 L Arginine 10 Polyvinylpyrrolidone 10 Microcrystalline Cellulose 40 Magnesium Stearate 2 Talc 2 Hydroxypropylmethylcellulose 6 PEG 6000 2 Titanium Dioxide 1 Eudragit L100 6 Acetylated Monoglycerides 1 STOP Layer Microcrystalline Cellulose 17 Starch maize 6 Crospovidone 2 Polyvinylpyrrolidone 3 OCA Layer UDCA Core Obeticholic acid 5 Microcrystalline Cellulose 20 Sodium Starch Glycollate 2 Colloidal Silicon Dioxide 1 Magnesium Stearate 2 Gastric acid soluble coating Opadry (Ready Film coating material) 12

Example 52

[0361]

TABLE-US-00027 Unit Composition UDCA + OCA combination Strength 200 + 10 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 200 Sodium Maleate 20 Polyvinylpyrrolidone 20 Microcrystalline Cellulose 80 Magnesium Stearate 4 Talc 4 Ethyl Cellulose 12 PEG 6000 4 Titanium Dioxide 2 Eudragit L100 12 Acetylated Monoglycerides 2 STOP Layer Microcrystalline Cellulose 30 Starch maize 10 Crospovidone 4 Polyvinylpyrrolidone 6 OCA Layer Obeticholic acid 20 Microcrystalline Cellulose 36 Sodium Starch Glycollate 4 Colloidal Silicon Dioxide 2 Magnesium Stearate 4 Gastric acid soluble coating Opadry (Ready Film coating material) 24

Example 53

[0362]

TABLE-US-00028 Unit Composition UDCA + OCA combination Strength 100 + 5 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 100 Disodium Phosphate 10 Polyvinylpyrrolidone 10 Microcrystalline Cellulose 40 Magnesium Stearate 2 Talc 2 Hydroxy Ethyl Cellulose 6 PEG 6000 2 Titanium Dioxide 1 Eudragit L55D (EQ to dried solid 6 content) Acetylated Monoglycerides 1 STOP Layer Microcrystalline Cellulose 17 Starch maize 6 Crospovidone 2 Polyvinylpyrrolidone 3 OCA Layer Obeticholic acid 5 Microcrystalline Cellulose 20 Sodium Starch Glycollate 2 Colloidal Silicon Dioxide 1 Magnesium Stearate 2 Gastric acid soluble coating Opadry (Ready Film coating material) 12

Example 54

[0363]

TABLE-US-00029 Unit Composition UDCA + OCA combination Strength 250 + 12.5 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 250 Sodium Bicarbobate 25 Polyvinylpyrrolidone 12 Microcrystalline Cellulose 75 Magnesium Stearate 5 Talc 3 Hydroxypropylmethylcellulose 7 PEG 6000 5 Titanium Dioxide 1 Hydroxypropylmethylcellulose Acetyl 12 Succinate Acetylated Monoglycerides 2 STOP Layer Microcrystalline Cellulose 3 Starch maize 1 Crospovidone 2 Polyvinylpyrrolidone 2 OCA Layer Obeticholic acid 12.5 Microcrystalline Cellulose 20 Sodium Starch Glycollate 4 Colloidal Silicon Dioxide 0.5 Magnesium Stearate 2 Gastric acid soluble coating Opadry (Ready Film coating material) 6

Example 55

[0364]

TABLE-US-00030 Unit Composition UDCA + OCA combination Strength 1000 + 50 Ingredient Quantity/Tablet (in mg) UDCA Core Ursodeoxycholic acid 1000 Sodium Bicarbobate 100 Polyvinylpyrrolidone 30 Microcrystalline Cellulose 300 Magnesium Stearate 20 Talc 13 Hydroxypropylmethylcellulose 28 PEG 6000 20 Titanium Dioxide 5 Hydroxypropylmethylcellulose Phthalate 50 Acetylated Monoglycerides 6 STOP Layer Microcrystalline Cellulose 30 Starch maize 15 Crospovidone 8 Polyvinylpyrrolidone 8 OCA Layer Obeticholic acid 50 Microcrystalline Cellulose 80 Sodium Starch Glycollate 8 Colloidal Silicon Dioxide 3 Magnesium Stearate 2 Gastric acid soluble coating Opadry (Ready Film coating material) 24

[0365] Examples of the production of multilayer tablets according to the invention are shown below in the Examples. The number of the components in the multilayer tablets is shown in the Tables provided in each example.

Example 56

[0366] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0367] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0368] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0369] The final granule blend is compressed into tablet cores.

[0370] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0371] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0372] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0373] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00031 Single tablet composition Ingredients UDCA + OCA, mg 100 + 1 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 100 L Arginine 10 Polyvinylpyrrolidone 10 Microcrystalline Cellulose 40 Magnesium Stearate 2 Talc 2 Hydroxypropylmethylcellulose 6 PEG 6000 2 Titanium Dioxide 1 Eudragit L100 6 Acetylated Monoglycerides 1 STOP Layer Microcrystalline Cellulose 17 Starch maize 6 Crospovidone 2 Polyvinylpyrrolidone 3 OCA Layer Obeticholic acid 1 Microcrystalline Cellulose 20 Sodium Starch Glycollate 2 Colloidal Silicon Dioxide 1 Magnesium Stearate 2 Gastric acid soluble layer Opadry (Ready Film coating material) 12

Example 57

[0374] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0375] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0376] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0377] The final granule blend is compressed into tablet cores.

[0378] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0379] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0380] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0381] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00032 Single tablet composition Ingredients UDCA + OCA, mg 150 + 1.5 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 150 L Arginine 20 Polyvinylpyrrolidone 20 Microcrystalline Cellulose 80 Magnesium Stearate 4 Talc 4 Hydroxypropylmethylcellulose 12 PEG 6000 4 Titanium Dioxide 2 Eudragit L100 12 Acetylated Monoglycerides 2 STOP Layer Microcrystalline Cellulose 30 Starch maize 10 Crospovidone 4 Polyvinylpyrrolidone 6 OCA Layer Obeticholic acid 1.5 Microcrystalline Cellulose 25 Sodium Starch Glycollate 4 Colloidal Silicon Dioxide 2 Magnesium Stearate 4 Gastric acid soluble layer Opadry (Ready Film coating material) 28

Example 58

[0382] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0383] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0384] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0385] The final granule blend is compressed into tablet cores.

[0386] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0387] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0388] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0389] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00033 Single tablet composition Ingredients UDCA + OCA, mg 200 + 2.5 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 200 L Arginine 30 Polyvinylpyrrolidone 25 Microcrystalline Cellulose 150 Magnesium Stearate 10 Talc 6 Hydroxypropylmethylcellulose 15 PEG 6000 10 Titanium Dioxide 2 Eudragit L100 24 Acetylated Monoglycerides 4 STOP Layer Microcrystalline Cellulose 36 Starch maize 14 Crospovidone 6 Polyvinylpyrrolidone 6 OCA Layer Obeticholic acid 2.5 Microcrystalline Cellulose 30 Sodium Starch Glycollate 6 Colloidal Silicon Dioxide 4 Magnesium Stearate 6 Gastric acid soluble layer Opadry (Ready Film coating material) 36

Example 59

[0390] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0391] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0392] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0393] The final granule blend is compressed into tablet cores.

[0394] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0395] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0396] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0397] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00034 Single tablet composition Ingredients UDCA + OCA, mg 250 + 5 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 250 L Arginine 40 Polyvinylpyrrolidone 30 Microcrystalline Cellulose 300 Magnesium Stearate 20 Talc 12 Hydroxypropylmethylcellulose 28 PEG 6000 20 Titanium Dioxide 5 Eudragit L100 44 Acetylated Monoglycerides 6 STOP Layer Microcrystalline Cellulose 40 Starch maize 20 Crospovidone 8 Polyvinylpyrrolidone 8 OCA Layer Obeticholic acid 5 Microcrystalline Cellulose 40 Sodium Starch Glycollate 8 Colloidal Silicon Dioxide 4 Magnesium Stearate 8 Gastric acid soluble layer Opadry (Ready Film coating material) 44

Example 60

[0398] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0399] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0400] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0401] The final granule blend is compressed into tablet cores.

[0402] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0403] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0404] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0405] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00035 Single tablet composition Ingredients UDCA + OCA, mg 500 + 5 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 500 L Arginine 45 Polyvinylpyrrolidone 34 Microcrystalline Cellulose 350 Magnesium Stearate 20 Talc 12 Hydroxypropylmethylcellulose 28 PEG 6000 20 Titanium Dioxide 5 Eudragit L100 44 Acetylated Monoglycerides 6 STOP Layer Microcrystalline Cellulose 45 Starch maize 25 Crospovidone 8 Polyvinylpyrrolidone 8 OCA Layer Obeticholic acid 5 Microcrystalline Cellulose 60 Sodium Starch Glycollate 8 Colloidal Silicon Dioxide 8 Magnesium Stearate 8 Gastric acid soluble layer Opadry (Ready Film coating material) 48

Example 61

[0406] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0407] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0408] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0409] The final granule blend is compressed into tablet cores.

[0410] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0411] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0412] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0413] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00036 Single tablet composition Ingredients UDCA + OCA, mg 750 + 10 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 750 L Arginine 50 Polyvinylpyrrolidone 40 Microcrystalline Cellulose 400 Magnesium Stearate 25 Talc 17 Hydroxypropylmethylcellulose 35 PEG 6000 26 Titanium Dioxide 7 Eudragit L100 50 Acetylated Monoglycerides 10 STOP Layer Microcrystalline Cellulose 55 Starch maize 30 Crospovidone 10 Polyvinylpyrrolidone 10 OCA Layer Obeticholic acid 10 Microcrystalline Cellulose 80 Sodium Starch Glycollate 10 Colloidal Silicon Dioxide 10 Magnesium Stearate 10 Gastric acid soluble layer Opadry (Ready Film coating material) 60

Example 62

[0414] The amount of each component in one tablet is shown in the Table below. For the purpose of manufacturing one hundred tablets, the mass of each component was 100-fold the mass specified in the Table.

[0415] Ursodeoxycholic acid, L-arginine, 50% microcrystalline cellulose, 50% hydroxypropylmethylcellulose and holyvinylpyrrolidone are dry mixed and wet granulated in an appropriate granulator with sufficient purified water.

[0416] The wet granules are dried, milled, and blended with the remaining 50% microcrystalline cellulose, remaining 50% hydroxypropylmethylcellulose, talc, magnesium stearate, PEG 6000, titanium dioxide, Eudragit L100 and acetylated monoglycerides.

[0417] The final granule blend is compressed into tablet cores.

[0418] Microcrystalline cellulose, starch maize, crospovidone, and polyvinylpyrrolidone are added slowly to purified water and mixing is continued until the components are fully dispersed.

[0419] The dispersion is sprayed on to the tablet cores in a conventional coating pan until proper amount of STOP layer is deposited on the tablet cores.

[0420] Obeticholic acid is dissolved in purified water. After thorough mixing, microcrystalline cellulose, sodium starch glycollate, colloidal silicon dioxide, and magnesium stearate are added slowly, and mixing is continued until the components are fully dispersed. The suspension is sprayed on to the tablet cores covered with STOP layer in a conventional coating pan until the proper amount of OCA layer is deposited.

[0421] Opadry is added slowly to purified water and mixing is continued until Opadry is fully dispersed. The solution is sprayed on to the tablet cores covered with STOP layer and OCA layer in a conventional coating pan until proper amount of Opadry is deposited on the tablets. A sample of the tablets is tested for gastric resistance and the coating stopped if the tablets pass the test.

TABLE-US-00037 Single tablet composition Ingredients UDCA + OCA, mg 1000 + 10 Ingredient Qty/Tablet (in mg) UDCA Core Ursodeoxycholic acid 1000 L Arginine 70 Polyvinylpyrrolidone 50 Microcrystalline Cellulose 500 Magnesium Stearate 35 Talc 24 Hydroxypropylmethylcellulose 45 PEG 6000 40 Titanium Dioxide 10 Eudragit L100 70 Acetylated Monoglycerides 15 STOP Layer Microcrystalline Cellulose 70 Starch maize 40 Crospovidone 12 Polyvinylpyrrolidone 12 OCA Layer Obeticholic acid 10 Microcrystalline Cellulose 100 Sodium Starch Glycollate 20 Colloidal Silicon Dioxide 15 Magnesium Stearate 15 Gastric acid soluble layer Opadry (Ready Film coating material) 80

[0422] In order for the patient to receive his daily dose of UDCA and OCA, he can take, for example, several tablets per day, indicated in examples 56-60 (for example, three or four tablets from example 2), or one tablet indicated in examples 61-62.

[0423] The dissolution data for one of the representatives of disclosed solid dosage forms is mentioned below (Table 17) for both OCA and UDCA actives in the given media:

TABLE-US-00038 TABLE 17 Values of the release rates of OCA and UDCA upon dissolution of the claimed solid dosage form, in % Percent Drug released in minutes Time (in mins.) 15 30 45 60 120 240 300 OCA 52 65 75 99 100 100 100 UDCA 0 0 0 0 23 48 99

[0424] A double-blind clinical study on the effectiveness of the combined pharmaceutical composition comprising UDCA and OCA compared to UDCA was conducted.

[0425] To study the effectiveness, the tablet embodiments No. 15 and No. 16 were used.

TABLE-US-00039 Embodiment 15 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release STOP-layer Layer No. 3 - obeticholic acid - 5 mg Outer layer - enteric coating Embodiment 16 Coated multilayer Core - ursodeoxycholic acid - tablet with modified 500 mg release and extended STOP-layer release of UDCA Layer No. 3 - obeticholic acid - 10 mg Outer layer - film non-enteric coating

[0426] A study on the effectiveness of the various treatment regimens for the patients with PBC was conducted. A total of three treatment regimens were used in the study: [0427] 1) UDCA monocomposition at a dose of 15 mg/kg/day. [0428] 2) Combined pharmaceutical composition in the pharmaceutical form with the modified release and at a dose of UDCA 15 mg/kg+OCA 5 mg per day. [0429] 3) Combined pharmaceutical composition in the pharmaceutical form with the modified release and at a dose of UDCA 15 mg/kg+OCA 10 mg per day.

[0430] The studies were carried out for 12 months, during which the quality of the treatment was assessed by measuring biochemical and molecular biological markers (see above).

[0431] The evaluation of the effectiveness of the treatment was carried out on the basis of the Paris2 system, according to which the criteria for a satisfactory treatment are: [0432] ALT level≤3-fold upper limit of normal (ULN); [0433] AST level≤2-fold ULN; [0434] normal bilirubin level.

TABLE-US-00040 TABLE 18 Effectiveness of the PBC treatment using various regimens Number of Number of patients with patients with positive positive Total response to response to number of treatment treatment patients (number) (%) UDCA 15 mg/kg 68 40 58.82 UDCA 15 mg/kg + 65 56 86.15 OCA 5 mg UDCA 15 mg/kg + 63 58 92.06 OCA 10 mg

[0435] According to the data above, the treatment regimen using a combined pharmaceutical composition in the pharmaceutical form with the modified release and at the dose of UDCA 15 mg/kg+OCA 10 mg per day, was found to be the most effective (92.06% of the patients had a positive result of the treatment). In second place, the treatment regimen using the combined pharmaceutical composition in the pharmaceutical form with the modified release and at the dose of UDCA 15 mg/kg+OCA 5 mg per day (86.15% of the patients had a positive result of the treatment). The regimen with the use of UDCA monocomposition had the lowest effectiveness−58.82%.

[0436] The technical contribution of the present disclosure resides in the following. Due to the selection and use of the new group of the additional markers, the authors were able to study the molecular processes in the liver and divide the patients by stages of PBC to apply the necessary treatment regimen and increase the effectiveness of the treatment up to 87% by using the highly compliant pharmaceutical composition that can apply to all stages of PBC and comprises UDCA and OCA together. The competition of ursodeoxycholic and obeticholic acids was also determined. In order to prevent the competition for the transporters, the new solid dosage form with a structural solution, such as the multilayer solid dosage form for the separate release of two acids, was developed.

[0437] The given embodiments of the present disclosure in no way limit it but are given only for a better understanding of its essence.