DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
The instant application, filed November 7, 2022, is a national stage entry of PCT/US2021/031277, filed May 7, 2021, which claims priority to U.S. Provisional Application No. 63/022,066, filed May 8, 2020. Receipt is acknowledged of certified copies of papers required by 37 CFR § 1.55.
Preliminary Amendments
The preliminary amendments filed on November 7, 2022, November 8, 2022, and April 28, 2023, are acknowledged and entered. Claims 4-9, 18, 27-43, and 45-94 are canceled. Claims 1-3, 10-17, and 19-26 are amended.
Information Disclosure Statement
The Information Disclosure Statements filed on February 7, 2023 (two filed on this date), April 28, 2023, and October 20, 2025, are acknowledged and found to be in compliance with the provisions of 37 CFR § 1.97. Accordingly, the Information Disclosure Statements have been considered.
Restriction/Election
Claims 1-3, 10-17, 19-26, and 44 are subject to a restriction and election requirement dated August 19, 2025. Applicants’ election without traverse of Group I (claims 1, 10-17, and 19-26) in the reply filed on October 20, 2025 is acknowledged.
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The election without traverse of the following species are also acknowledged:
The species of vitamin-D receptor (“VDR”) agonist:
A VDR agonist that induces GLP-1 secretion from a target cell as in claim 11.
The species of agent:
Lithocholic acid (“LCA”) - including derivative thereof, as in claim 16.
The Specification provides the following chemical name and structure for LCA:
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Specification at 57, paragraph [00206].
The species of disease for treatment:
Diabetes, as in claim 1.
Status of Claims
Claims 1-3, 10-17, 19-26, and 44 are pending in the instant application. Claims 2, 3, 12, 22 and 44 are withdrawn from further consideration pursuant to 37 CFR § 1.142(b), as being drawn to a non-elected invention and species. Therefore, claims 1, 10, 11, 13-17, 19-21, and 23-26 read on an elected invention and species and are therefore under consideration in the instant application.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 16, 20, 25, and 26 rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 16 is Indefinite
Claim 16 recites the method of claim 1, “wherein the agent is lithocholic acid (LCA), or a derivative of LCA, or a pharmaceutically acceptable salt thereof” (emphasis added). The scope claim is unclear because the meaning of the limitation “a derivative of LCA” is unclear.
The Specification defines a “derivative” as follows:
The term “derivative” as used herein means any chemical, conservative substitution, or structural modification of an agent. The derivative can improve characteristics of the agent or small molecule such as pharmacodynamics, pharmacokinetics, absorption, distribution, delivery, targeting to a specific receptor, or efficacy. For example, for a small molecule, the derivative can consist essentially of at least one chemical modification to about ten modifications. The derivative can also be the corresponding salt of the agent. The derivative can be the pro-drug of the small molecule as provided herein.
Specification at 53, paragraph [00195].
When the special definition of a “derivative” is applied to the term “a derivative” in claim 16, claim 16 recites (emphasis added):
A method for treating diabetes, obesity, or an inflammatory disease in a subject in need thereof, the method comprising administering to the subject an agent that increases levels or activity of a sulfotransferase in the subject, wherein the agent is lithocholic acid (LCA), or any chemical, conservative substitution, or structural modification of LCA, or a pharmaceutically acceptable salt thereof.
As an initial matter, the definition of a “derivative” lacks the necessary clarity required for distinctly claiming the subject matter of claim 16 because it is unclear if “chemical” modifies “conservative substitution”, or if “chemical” is one of three types of possible modifications of LCA (viz. “chemical modification”, “conservative substitution modification”, or “structural modification” of LCA).
In either reading, the Specification offers no guidance for how to interpret the phrases “conservative substitution” and “structural modification” of an agent, to the extent that “a derivative” encompasses any chemical. A “conservative substitution” implies the exchange of one chemical substituent for another, or put another way, one modification. Similarly, a “structural modification” implies one modification, and that the modification is somehow not a “conservative substitution”. However, as the Specification explains in the excerpted paragraph above, for a small molecule such as LCA, a derivative may include “at least one chemical modification to about ten modifications” (emphases added). As such, Applicants may intend the scope of any “conservative substitution” or “structural modification” of an agent to include at least up “to about ten modifications”, and perhaps more. Therefore, it is unclear if the scope of the limitation “a derivative of LCA” encompasses any chemical.1 As a result, one of ordinary skill in the art would not be apprised of the scope of the claim because the chemicals encompassed by the limitation “a derivative of LCA” are unclear. Accordingly, claim 16 is indefinite.
Applicants are required to amend the claim to clarify the scope of the limitation “a derivative of LCA”. For example, an amendment that specifies the chemicals which are derivatives of LCA, would provide necessary clarity and resolve the issue. The examiner acknowledges that examples of “Derivatives” of certain molecules are exemplified in the instant Figures 22 and 23, but notes that none of the structures appear to show “Derivatives” of “lithocholic acid”. Figures 22 and 23 are reproduced below.
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Specification at Fig. 22.
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Specification at Fig. 23.
Claim 20 is Indefinite
Claim 20 depends upon claim 19, which depends upon claim 1. When claim 20 is combined with the dependent claim 19 and the independent claim 1, it recites (emphases added):
A method for treating diabetes, obesity, or an inflammatory disease in a subject in need thereof, the method comprising administering to the subject an agent that increases levels or activity of a sulfotransferase in the subject,
wherein the agent is formulated with a pharmaceutical composition,
wherein the pharmaceutical composition is formulated to restrict delivery of an agent to the gastrointestinal tract of the subject.
The term “an agent” in the final wherein clause lacks clear antecedent basis and renders the scope of the claim unclear. In particular, it is unclear if the “an agent” recited in the final wherein clause is intended to refer to:
the same agent administered to a subject “that increases level or activity of a sulfotransferase in the subject” and “is formulated with a pharmaceutical composition”, or
a second agent whose delivery is being restricted by the pharmaceutical composition.
Because the term “an agent”, given its broadest reasonable interpretation, is such that a person of ordinary skill in the art would read it with more than one reasonable interpretation, the scope of claim 20 is not reasonably ascertainable.2 See MPEP 2173.02, subsection I (“For example, if the language of a claim, given its broadest reasonable interpretation, is such that a person of ordinary skill in the relevant art would read it with more than one reasonable interpretation, then a rejection under 35 U.S.C. 112(b) or pre-AIA 35 U.S.C. 112, second paragraph is appropriate.”). Therefore, claim 20 is indefinite.
Applicants are required to amend the claim to clarify the relationship between the various recitations of “agent”. For example, an amendment that specifies the pharmaceutical composition is formulated to restrict delivery of “the agent”, or that specifies the pharmaceutical composition is formulated to restrict delivery of “a second agent”, would provide clear antecedent basis and resolve the multiple interpretation issue.
Claim 25 is Indefinite
Claim 25 depends upon claim 1 and recites the that “the mammal is a human” (emphasis added). There is insufficient antecedent basis for the “mammal” limitation in the claim, and it is unclear if the “mammal” is the subject that has a disease being treated in the independent claim.
Claim 1 recites a method of treating a disease “in a subject”. However, claim 24 depends upon claim 1, and recites that “the subject” is a mammal. If all subjects are mammals, then claim 24 would be duplicative of claim 1, and claim 24 would be rejected under 35 U.S.C. 112(d) for failing further limit the subject matter of claim 1. Therefore, it is unclear if applicants intend “the mammal” limitation of claim 25 to further limit the “subject” recited in claim 1, or if in fact all subjects are mammals. Accordingly, the recitation of “the mammal” limitation is indefinite.
Applicants are required to amend the claim to provide sufficient antecedent basis for the “mammal” limitation. For example, an amendment that specifies that claim 25 depends upon claim 24, would provide clear antecedent basis for the “mammal” limitation.
Claim 26 is Indefinite
Claim 26 recites that “the target cell is a hepatocyte, enteroendocrine cell, an epithelial cell, an L-cell, or a neuron” (emphasis added). It is unclear the function of the “target cell” in the context of the claim.
For example, it is not clear whether the method of claim 26 requires that:
the agent be delivered selectively to the “target cell”,
the agent exert its primary pharmacological effect in the “target cell”,
the increased levels or activity of a sulfotransferase in the subject occur only in the “target cell”, or
some combination of the above.
For example, the Specification uses the specific phrase “target cell” twice. See Specification at 58, paragraph [00212] (“In another embodiment of any of the aspects, the TGR5 agonist induces GLP-1 secretion from a target cell. In some embodiments of any of the aspects, the target cell is an enteroendocrine cell, an epithelial cell, an L-cell, or a neuron.”) (emphases added). The specification further explains that hepatocytes may be involved in the sulfation of bile acids. See Specification at 57, section “Molecular Targets for the treatment of Diabetes, Obesity, and Inflammatory Diseases”, and in particular paragraph [00219] (“For example, activation of SULT2A, e.g., SULT2A1 results in the sulfation of bile acids in a cell (e.g. hepatocytes).”). Therefore, it is unclear the function of “the target cell” in the context of claim 26 given the separate molecular pathways that the instantly claimed agents may increase levels or activity of a sulfotransferase in the subject. Accordingly, claim 26 is indefinite.
Applicants are required to amend the claim to provide clarity for the function of “the target cell” limitation. For example, an amendment that specifies that claim 26 depends upon claim 11, would provide clarity for the function of “the target cell” limitation.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Background – Sulfotransferases
The instant claims are generally directed to method of treating diabetes in a subject by altering the levels or activity of a sulfotransferase. The following provides a background on sulfotransferases in the context of bile acids.
By way of background, reference is made to Dawson, Paul A., and Kenneth DR Setchell. "Will the real bile acid sulfotransferase please stand up? Identification of Sult2a8 as a major hepatic bile acid sulfonating enzyme in mice", Journal of lipid research, vol. 58, no. 6, pp. 1033-1035 (2017), hereinafter “Dawson 2017”. The teachings of Dawson 2017 are not incorporated into the body of the following rejection unless expressly stated.
Dawson 2017 generally discloses the role of sulfotransferases in the context of bile acids.
Dawson 2017 explains that bile acids undergo sulfonation in the liver and other tissues which alters their “physiochemical, pharmacokinetic, and singling properties” and “reduces their cytotoxicity.” See Dawson 2017 at 1033.
This year marks the 50th anniversary of the publication by Robert Palmer (1, 2) recognizing the formation of bile acid sulfates as a mechanism for bile acid elimination in humans. Like steroids, bile acids undergo sulfonation in liver and other tissues [reviewed by Alnouti in (3)]. This important phase II detoxification reaction transfers a sulfonate group (SO3−) from the universal donor, 3′-phosphoadenosine 5′-phosphosulfate (PAPS), to a hydroxyl, amine, or carboxylic acid group of a substrate. In humans, almost half of the lithocholic acid (LCA) secreted into bile is present in divalent, sulfated, and amidated form (4), and bile acid sulfates comprise a large proportion of the bile acids excreted in urine (5). Sulfonation is not specific to LCA because sulfated forms of cholic acid, deoxycholic acid, chenodeoxycholic acid, and ursodeoxycholic acid are also found in serum and urine, usually as double-conjugates with glycine and taurine in the side-chain (6). Notably, the levels of bile acid sulfates are elevated in physiological and pathophysiological cholestasis (7, 8). Sulfonation significantly alters the physicochemical, pharmacokinetic, and signaling properties of bile acids, reducing their cytotoxicity. For example, polar bile acid sulfates are choleretic, and sulfation promotes rapid elimination of bile acids by limiting their intestinal reabsorption and by efflux of divalent bile acid sulfates from liver into blood to increase their urinary excretion. In general, there is an inverse relationship between hydroxylation and sulfation whereby addition of hydroxyl groups prevents sulfation. In contrast to humans, hydroxylation rather than sulfation is thought to be the more dominant pathway for bile acid detoxification in mice (3, 9). However, sulfonation may still have a role under certain pathophysiological conditions in mice (10, 11). Another species difference between humans and mice is the position of the sulfate moiety on the steroid nucleus. Bile acids are primarily mono-sulfated in humans (although traces of disulfates are found in urine, possibly through renal sulfation) and in mice. However, the location of the sulfate group on the bile acid steroid nucleus differs, preferentially occurring at the C-3 position in humans and at the C-7 position in mice (with the exception of LCA, which is a minor bile acid in mice) (3, 6, 12). This is important because the sulfate moiety's position has a profound effect on the physiological behavior of bile acids. Unlike bile acids sulfated at the C-3 position, C-7-sulfates are resistant to hydrolysis and metabolism by the intestinal microbiota (13), thereby blocking their intestinal reabsorption and potentially providing cytoprotection to the intestinal mucosa (14).
Dawson 2017 at 1033 (emphases added).
Sulfonation is carried out by members of the superfamily of cytosolic sulfotransferases (SULTs). In humans, there are 13 SULT genes, which are distributed between 4 families (15). SULT2A1, the lone SULT2A gene in humans, appears to be the major human SULT enzyme capable of sulfonating bile acids at the 3α-OH position (15, 16, 17). In mice, there are 21 SULT genes, and these are subdivided between 6 families. Whereas there is only one human SULT2A gene, seven (and now eight) Sult2a genes are present in mice (15, 18). Most mouse studies have focused on Sult2a1 or Sult2a1/Sult2a2 as the putative hepatic bile acid sulfotransferase. However, the significant female predominance of hepatic Sult2a1 expression and dehydroepiandrosterone (DHEA) sulfotransferase activity in mice (19, 20) is not associated with a greater abundance of bile acid sulfates in urine or tissue (12). These observations as well as the plethora of mouse Sult2a genes raise questions as to which mouse ortholog(s) of human SULT2A1 are responsible for this important reaction and whether expression of Sult2a1 mRNA/protein can be equated with hepatic synthesis of bile acid sulfates. In the present issue of the Journal of Lipid Research, Feng et al. (21) identify a novel Sult2a isoform that begins to answer these questions.
Dawson 2017 at 1033 (emphases added).
Figure 1 of Dawson 2017 depicts the general mechanism for sulfonation of bile acids in humans and mice by sulfotransferases, reproduced below:
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Dawson 2017 at 1034.
Claims 1, 10, 11, 13-17, 19-21, and 23-26 are Obvious over WO’961 in view Ferrell 2019 and Chatterjee 2005
Claims 1, 10, 11, 13-17, 19-21, and 23-26 are rejected under 35 U.S.C. 103 as being unpatentable over Evans et al, “USE OF VITAMIN D RECEPTOR AGONISTS AND PRECURSORS TO TREAT FIBROSIS”, International Publication No. WO 2009/061961 A1, published May 14, 2009, hereinafter “WO’961”, in view of Ferrell, Jessica M. and Chiang, John Y. L. “Understanding Bile Acid Signaling in Diabetes: From Pathophysiology to Therapeutic Targets”, Diabetes & Metabolism Journal, vol. 43, no. 3, pp. 257-272 (2019), hereinafter “Ferrell 2019”, and Chatterjee, Bandana, Ibtissam Echchgadda, and Chung Seog Song. "Vitamin D receptor regulation of the steroid/bile acid sulfotransferase SULT2A1", Methods in enzymology, vol. 400 pp. 165-191 (2005), hereinafter “Chatterjee 2005”.
The instant claims are generally directed to a method of treating diabetes in a subject comprising administering lithocholic that increases the levels or activity of a sulfotransferase in the subject. In some of the claims and in certain embodiments, lithocholic acid acts as a vitamin D receptor (“VDR”) agonist that induces GLP-1 secretion from a target cell.
Generally, Applicants explain that:
Selective transport of the microbial metabolite lithocholic acid (LCA) from the gut to the liver after bariatric surgery activates hepatic vitamin D receptor (VDR), thereby inducing expression of bile acid sulfotransferase SULT2A, which produces the antidiabetic molecule cholic acid 7-sulfate (CA7S).
Specification at 2, paragraph [0006], and that
cholic acid 7-sulfate is a TGR5 agonist and induces GLP-1 secretion in vitro.
Specification at 18, paragraph [0065].
However, WO’961 teaches methods of treating fibrosis, a common complication of diabetes, comprising administering a VDR agonist, such as LCA. WO’961 further explains that LCA induces SULT2A expression.
WO’961
WO’961 exemplifies methods of treating fibrosis in a subject comprising administering vitamin D receptor agonists.
Described herein are methods of treating fibrosis that are based on the unexpected discovery that a specialized subset of cells within the liver responds to compounds that bind to or activate the vitamin D receptor (VDR, NRlIl) to influence the processes of liver injury, inflammation and fibrogenesis. Cells that express and respond to the VDR in liver include, but are not limited to hepatic stellate cells (HSCs), myofibroblasts, Kupffer cells (KCs), and sinusoidal endothelial cells (SECs). These cells types are frequently referred to as hepatic non-parenchymal cells (NPCs). In addition, there are similar specialized cells within the pancreas and kidney that respond in a similar manner, including but not limited to pancreatic stellate cells and renal mesangial cells.
Thus, one embodiment of the disclosure is a method of treating fibrosis in a subject. The method can include administering a therapeutically effective amount of vitamin D receptor agonist (such as lα,25 dihydroxyvitamin D3, (1,25-(OH)2- D3) or a precursor thereof, a vitamin D analog, a vitamin D receptor ligand, or a vitamin D receptor agonist precursor), to a subject having a fibrosis or at risk for developing fibrosis, thereby treating the fibrosis.
WO’961 at 2-3 (emphases added).
WO’961 explains that diabetes contributes to the high prevalence of end stage renal disease (“ESRD”), and that ESRD is characterized by significant renal fibrosis:
Almost all forms of end stage renal disease (ESRD) are characterized by significant renal fibrosis. A number of cardiovascular diseases, the aging population, and diabetes contribute to the high prevalence of ESRD. Two-thirds of ESRD patients are treated by frequent (2-3 times weekly) and long dialysis sessions and one-third is treated by kidney transplantation. In Europe, kidney replacement therapy is consuming 2% if the healthcare budget for only 0.1% of the population being treated.
WO’961 at 2 (emphasis added).
WO’961 further explains that hepatic fibrosis and pancreatic fibrosis are driven by inflammatory changes:3
Hepatic fibrosis, the accumulation of abnormal extracellular matrix (ECM) proteins and a resultant loss of liver function, is an accompaniment of an inflammation-driven wound healing process triggered by chronic liver injury. The main causes of liver injury leading to fibrosis in Western societies include chronic hepatitis C virus (HCV) infection, alcohol abuse, chronic hepatitis B (HBV) infection, iron overload as occurs in hereditary hemochromatosis, and increasingly, non-alcoholic steatohepatitis (NASH). The inflammatory process ensuing from hepatic injury triggers a variety of cellular responses including cell repair, hepatocyte regeneration, increased extracellular matrix turnover, and ultimately in some patients significant fibrosis. Progressive fibrosis of the liver eventually can result in cirrhosis, portal hypertension and hepatocelluar carcinoma.
WO’961 at 1, and
Fibrosis of the pancreas is caused by such processes as necrosis/apoptosis, inflammation, and duct obstruction. The initial event that induces fibrogenesis in the pancreas is an injury that may involve the interstitial mesenchymal cells, the duct cells and/or the acinar cells. Damage to any one of these tissue compartments of the pancreas is associated with cytokine-triggered transformation of resident fibroblasts/pancreatic stellate cells into myofibroblasts and the subsequent production and deposition of extracellular matrix. Depending on the site of injury in the pancreas and the involved tissue compartment, predominantly inter(peri)lobular fibrosis (as in alcoholic chronic pancreatitis), periductal fibrosis (as in hereditary pancreatitis), periductal and interlobular fibrosis (as in autoimmune pancreatitis) or diffuse inter- and intralobular fibrosis (as in obstructive chronic pancreatitis) develops. Given the foregoing, it would be desirable to have methods of treating, preventing, and ameliorating fibrosis, such as fibrosis of the liver, kidney, or pancreas.
WO’961 at 2.
WO’961 explains that treatment of hepatic non-parenchymal cells (“NPCs”)4 with VDR agonists slows, prevents, or reverses inflammatory and fibrotic changes to the liver:
Treatment of hepatic NPCs with VDR agonists has profound effects on gene expression in NPCs. For example, when HSCs are cultured on plastic, they undergo a process called "activation," wherein they change phenotype from a retinol- and lipid-rich cell into an extracellular matrix-producing cell that is ultimately responsible for the production of scarring within the liver (fibrogenesis). VDR ligands prevent or retard this activation process, and reverse the process in some embodiments. Moreover, treatment of HSCs with VDR ligands markedly attenuates pro-inflammatory and pro-fibrotic gene expression induced by treating HSCs with either bacterial lipopolysaccharide endotoxin (LPS) or transforming growth factor beta 1 (TGF-βl). In particular, VDR ligands attenuate or abrogate LPS-induced pro-inflammatory chemokine production and TGF-β-induced pro- fibrotic collagen production by HSCs (Table 1). LPS is a potent activator of the innate immune system while TGF-β is a family of three proteins that regulate differentiation, proliferation and many other functions in a wide range of cell types. Thus, VDR ligands and other VDR agonists play a therapeutic role in the prevention of liver injury, inflammation, and fibrogenesis in persons with liver diseases, including but not limited to chronic viral hepatitis (Hepatitis B and Hepatitis C infection), alcohol-induced liver disease, non-alcoholic steatohepatitis, autoimmune liver diseases, and genetic liver diseases, such as hereditary hemochromatosis, alpharantitrysin deficiency and Wilson's disease.
WO’961 at 20.
WO’961 teaches that 1) VDR is highly expressed in NPCs, 2) lithocholic acid (“LCA”) activates VDR, and 3) LCA induces SULT2A1 expression:
Despite its relatively high expression level in NPCs, the role of VDR in these cells was unknown prior to this disclosure. VDR possesses the common nuclear receptor structure, for instance is comprised of an N-terminal activation domain, a DNA-binding region (DBD) with two zinc finger domains, a hinge region and a ligand-binding domain (LBD). VDR activated gene transcription requires initial nuclear translocation via importin-α, heterodimerization with RXR, (Yasmin et al, 2005. J Biol Chem., 280(48):40152-60), and binding to response elements present in target genes. VDR regulates genes associated with the maintenance of calcium and phosphate homeostasis in the intestine and kidney. The signal initiated by VDR/RXR heterodimers is modulated by the association of co-activating or co-repressing proteins and also depends on other signaling partners in the nuclear compartment (Ebert et ah, 2006. MoI Cell Endocrinol., 248(1-2): 149-59). The VDR/RXR heterodimer is non-permissive, in that the presence or absence of RXR ligands does not affect VDR responses (Shulman et ah, 2004. Cell, 116(3):417-29). Until recently, the only known physiological ligand for VDR was calcitriol. However, specific bile acids such as LCA and some derivatives (LCA-acetate, LC A- formate, 3-keto LCA) may activate VDR. These bile acid VDR agonists have been shown to induce SULT2A1 expression, a sulfo-conjugating phase II enzyme in intestinal mucosa, which may provide a key defense response of the intestine against the toxic and carcinogenic effects of bile acids (Chatterjee et ah, 2005. Methods Enzymoh, 400: 165-91).
WO’961 at 22-23 (emphases added).
WO’961 explains that the therapeutic benefit of VDR agonism in the liver was overlooked because hepatocytes usually exhibit very low levels of VDR:
It was previously thought that the liver lacked VDR expression because hepatocytes, the most abundant cell population in liver, usually exhibit very low levels of the receptor: the total level of VDR in rat liver is 1, 300-fold lower than in intestine. It is possible that the increase in intracellular Ca2+ levels observed in rat hepatocytes in response to 1,25-(OH)2-D3 may be due to an unrelated membrane receptor or an indirect mechanism rather than VDR-mediated signaling (Mailhot et ah, 2000. Endocrinology., 141 :891-900). However, 1,25-(OH)2-D3 has a significant effect on liver cell physiology during the compensatory growth process following the partial hepatectomy in the rat (Segura et ah, 1999. Histochem Cell Biol., 112(2): 163-7; Gascon-Barre et α/., 1994. J Clin Invest, 93(5):2159-67). Thus, VDR expression was examined in freshly isolated hepatic NPC populations. Surprisingly, it was shown herein that VDR is abundantly expressed in HSCs, KCs and SECs isolated from normal rat livers and the VDR in these cells is fully functional as determined by the VDR-dependent induction of CYP24A1 expression by lα,25-(OH)2-D3.
WO’961 at 23 (emphases added).
Therefore, WO’961 discloses 1) a method of treating diabetes in a subject (by treating or reducing the risk of fibrosis) comprising administering a VDR agonist, 2) discloses that LCA is a VDR agonist, and 3) that LCA induces expression of a sulfotransferase (SULT2A1).
While WO’961 teaches every limitation of claim 1, the following references provide motivation to adapt the method of treating diabetes (by treating or reducing the risk of fibrosis) disclosed in WO’961 to more generally expand upon the known therapeutic benefits that VDR agonism provides for diabetic subjects.
For example,
Ferrell 2019 explains that LCA is known to be a potent endogenous TGR5 agonist, and that activation of TGR5 stimulates the release of GLP-1 from enteroendocrine L-cells to stimulate insulin secretion from β-cells and increase insulin sensitivity.
Chatterjee 2005 explains that LCA is known activate VDR, which in turn causes the activation of SULT2A to detoxify LCA.
Ferrell 2019
Ferrell 2019 discloses that the Takeda G protein-coupled receptor 5 (“TGR5”) is agonized by LCA, and that TGR5 activation simulates release of GLP-1.
In the liver, TGR5 is expressed in sinusoidal endothelial cells, Kupffer cells (hepatic resident macrophages), stellate cells, and biliary epithelial cells in bile ducts, but not in hepatocytes [39-41], and the secondary bile acids LCA and DCA are potent en-dogenous TGR5 agonists (LCA>DCA>CDCA>CA). TGR5 is expressed in the epithelium of human gallbladder and con-trols gallbladder refiling [42]. TGR5 also plays a key role in bile acid metabolism and fasting-induced hepatic steatosis [43]. In the colon, TGR5 mediates bile acid-induced gastrointestinal motility, transit time and defecation [44]. In the intestine and macrophage, activation of TGR5 protects against inflamma-tion [45]. TGR5 also plays a critical role in the control of glu-cose homeostasis [46]. Activation of TGR5 stimulates the re-lease of glucagon-like peptide-1 (GLP-1) from enteroendo-crine L-cells to stimulate insulin secretion from β-cells and in-crease insulin sensitivity. In adipose tissue, activation of TGR5 induces thyroid hormone deiodinase type 2 (DIO2), which converts thyroid hormone thyroxine (T4) to triiodothyronine (T3) to stimulate energy metabolism and white adipose tissue browning (Fig. 2). CDCA increases brown adipose tissue in humans, likely through TGR5-mediated increase of uncou-pling protein and DIO2 expression [47]. TGR5 knockout mice are protected from cholesterol gallstone disease and high fat diet (HFD) induced obesity [48,49]. FXR and TGR5 are co-ex-pressed in L-cells, and activation of intestinal FXR stimulates TGR5 gene transcription via an FXR response element located in the TGR5 gene promoter; this crosstalk stimulates GLP-1 secretion [50]. It is therefore likely that some of the reported FXR effects on glucose metabolism may be due to TGR5 sig-naling in the gut.
Ferrell 2019 at 261 (emphases added).
Chatterjee 2005
Chatterjee 2005 explains that endogenous SULT2A1 mRNA and protein expression is induced by vitamin D:
Endogenous SULT2A1 mRNA and protein expression is induced by vitamin D (Echchgadda et al., 2004a). Figure 2 shows an example of this induction in liver cells and in the mouse liver. Treatment of HepG2 hepatoma cells with vitamin D (1α,25‐dihydroxy vitamin D3) for 24 h induced SULT2A1 mRNAs ∼6‐fold in a RT‐PCR assay (Fig. 2A); in mice injected with vitamin D, Sult2A1 mRNAs in the liver increased ∼3‐fold in a Northern blot assay (Fig. 2B). This induction reflected activation of the corresponding promoter (Fig. 3). In Caco‐2 intestinal cells, vitamin D induced the human promoter (from −1102 to 10) ∼6‐fold and the mouse promoter (from −292 to 42) ∼4.5‐fold; similar induction of the promoter has been observed in HepG2 hepatoma cells. Vitamin D induction of the human CYP3A4 promoter (from the CYP3A‐Luc construct) is also shown (Fig. 3A). This construct contains a proximal (at −173) and a distal (at −7715) VDR‐binding element (Drocourt 2002, Makishima 2002) in the CYP3A4 promoter.
Chatterjee at 174-175.
Figure 2 of Chatterjee 2005 is reproduced below:
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Chatterjee 2005 at 174, Figure 2.
Chatterjee 2005 explains that LCA and its derivatives agonize VDR and induce SULT2A1:
Secondary bile acid lithocholic acid (LCA) and its derivatives (LCA‐acetate, LCA‐formate, 3‐keto LCA) are known to serve as sensitive VDR agonists and to induce VDR target genes (Adachi 2005, Makishima 2002). Figure 3B shows induction of the SULT2A1 promoter by LCA‐acetate. The LCA‐mediated induction of SULT2A1 may be a key defense response of the intestinal and hepatobiliary tissues against the toxic/carcinogenic effects of bile acids.
Chatterjee 2005 at 175.
Chatterjee 2005 explains that the LCA-mediated induction of SULT2A1 may have emerged to protected from the toxicity of disrupted bile acid/sterol homeostasis caused by endogenous or exogenous factors:
LCA and LCA derivatives are agonist ligands of VDR and are able to induce CYP2, CYP3, and several other VDR target genes at micro molar concentrations (Adachi et al., 2005). The SULT2A1 promoter is also induced by LCA‐acetate (Fig. 3B). We predict that VDR takes a central role in LCA‐mediated induction of the SULT2A1 gene, even though PXR and FXR, the two NRs that are also activated by LCA, can induce SULT2A1 (Song et al., 2001; Sonoda et al., 2002). This prediction is made on the ground that LCA is an FXR antagonist (Yu et al., 2002) and that the functional LCA concentration for PXR activation is about 10‐fold higher than that needed to activate VDR (Makishima et al., 2002). In conclusion, a role for the VDR pathway in the basal and induced expression of SULT2A1, a phase II transferase, may have emerged to protect cells of the first‐pass tissues from the adverse effects of disrupted bile acid/sterol homeostasis caused by endogenous or exogenous factors.
Chatterjee 2005 at 187-188 (emphases added).
Claim 1 was Obvious at the Time of Filing
One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in developing a method of treating diabetes in a subject in need thereof comprising administering an agent that increases the levels of a sulfotransferase in the subject because WO’961 discloses that 1) fibrosis, a common complication of diabetes, may be treated by administering a VDR agonist, 2) LCA is a VDR agonist, and 3) LCA induces the expression of SULT2A1, a sulfotransferase. On having ordinary skill in the art would be further motivated to develop a method of treating diabetes comprising administering LCA because:
Ferrell 2019 explains that LCA is known to be a potent endogenous TGR5 agonist, and that TGR5 activation stimulates the release of GLP-1 from enteroendocrine L-cells to stimulate insulin secretion from β-cells and increase insulin sensitivity, and
Chatterjee 2005 explains that LCA and its derivatives serve as sensitive VDR agonists, and that the LCA-mediated induction of SULT2A1 may protect cells of the first‐pass tissues from the adverse effects of disrupted bile acid/sterol homeostasis caused by endogenous or exogenous factors.
Therefore, claim 1 was obvious at the time of filing.
Claim 10 was Obvious at the Time of Filing
Claim 10 recites the method of claim 1, wherein the agent is a VDR agonist.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
The above rejection for claim 1 explains that the claimed method was obvious when the agent is LCA, a VDR agonist.
Therefore, claim 10 was obvious for the same reasons claim 1 was obvious.
Claim 11 was Obvious at the Time of Filing
Claim 11 recites the method of claim 10, wherein the VDR agonist induces GLP-1 secretion from a target cell.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
Ferrell 2019 explains that LCA is a potent TGR5 agonist and that TGR5 activation simulates GLP-1 release from enteroendocrine L-cells.
Therefore, claim 11 was obvious for the same reasons claim 1 was obvious.
Claims 13-17 were Obvious at the Time of Filing
Claims 13-17 recite different agents for use in the method of claim 1, wherein the agents are small molecules or LCA or its derivatives.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
The above rejection for claim 1 explains that the claimed method was obvious when the agent is LCA. LCA is a small molecule according to the Specification. See Specification at 52-53, paragraph [00193] (defining “small molecule” to include an organic molecule having a molecular weight less than about 10,000 grams per mole).
Further, while claim 16 is rejected as indefinite because the limitation “a derivative of LCA” lacks sufficient clarity, Chatterjee 2005 explains that LCA derivatives are also VDR agonists.
Therefore, claims 13-17 were obvious for the same reasons claim 1 was obvious.
Claim 19 was Obvious at the Time of Filing
Claim 19 recites the method of claim 1, wherein the agent is formulated with a pharmaceutical composition.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
WO’961 discloses pharmaceutical compositions of the VDR agonists. See, e.g., WO’961 at 37:
The present disclosure includes a treatment for fibrosis, for instance hepatic, renal or pancreatic fibrosis, in a subject. The method includes administering vitamin D receptor agonists, such as lα,25(OH)2 D3, vitamin D precursors (for instance, 25-hydroxy-D3 (25-OH-D3) (calcidiol); vitamin D3 (cholecalciferol); or vitamin D2 (ergocalciferol)), vitamin D analogs, and vitamin D receptor agonists precursors to the subject in a pharmaceutically acceptable carrier and in an amount effective to inhibit (for example to relieve, cure, ameliorate, or prevent) the development, progression, or manifestation of fibrosis in the subject. The present disclosure also contemplates the administration of a therapeutic composition comprising more than one VDR agonist, as well as VDR agonists in combination with other therapies.
The vehicle in which the VDR agonist is delivered can include pharmaceutically acceptable compositions of the compounds, using methods well known to those with skill in the art. Any of the common carriers, such as sterile saline or glucose solution, can be utilized. The vehicle also can contain conventional pharmaceutical adjunct materials such as, for example, pharmaceutically acceptable salts to adjust the osmotic pressure, lipid carriers such as cyclodextrins, proteins such as serum albumin, hydrophilic agents such as methyl cellulose, detergents, buffers, preservatives and the like. A more complete explanation of parenteral pharmaceutical carriers can be found in Remington: The Science and Practice of Pharmacy (19th Edition, 1995) in chapter 95.
Embodiments of other pharmaceutical compositions can be prepared with conventional pharmaceutically acceptable carriers, adjuvants, and counter-ions, as would be known to those of skill in the art. The compositions in some embodiments are in the form of a unit dose in solid, semi-solid, and liquid dosage forms, such as tablets, pills, capsules, lozenges, powders, liquid solutions, or suspensions.
In some embodiments, sustained release of the pharmaceutical preparation that includes an effective amount of a VDR agonist is beneficial. Slow-release formulations are known to those of ordinary skill in the art. By way of example, sustained-release tablets can be formulated so that the active ingredient is embedded in a matrix of insoluble substance so that the dissolving drug emerges gradually through the holes in the matrix. In some formulations, the matrix physically swells to form a gel, so that the drug has first to dissolve in matrix, then exit through the outer surface.
WO’961 at 37-38.
One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in formulating the agent (i.e., the VDR agonist LCA) with a pharmaceutical composition because WO’961 explains that the VDR agonist may be administered in pharmaceutical compositions.
Therefore, claim 19 was obvious for the reasons given above, and for the same reasons claim 1 was obvious.
Claim 20 was Obvious at the Time of Filing
Claim 20 recites the method of claim 19, wherein the pharmaceutical composition is formulated to restrict delivery of an agent to the gastrointestinal tract of the subject.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
While claim 20 is rejected as indefinite because the limitation “an agent” lacks sufficient clarity, WO’961 explains that the VDR agonist, which may be formulated with a pharmaceutical composition as explained above in claim 19, may be delivered rectally or to target tissue:
It is not intended that the present disclosure be limited to a particular mode of administration. A variety of modes of administration are contemplated, including intravenously, intramuscularly, subcutaneously, intradermally, intraperitoneally, intrapleurally, intrathecally, orally, rectally, transdermally, by inhalation, and topically. In certain embodiments, the therapeutic compositions are administered via suppository, or in tablet or capsule formulations for oral delivery. In one embodiment, administration of the therapeutic compositions occurs at night. In another embodiment, multiple doses (e.g., 3 or 4) are provided in a 24 hour period. In a further embodiment, the administration of the therapeutic composition is by pulse intravenous therapy. In one example, the therapeutic compositions are administered via a transdermal patch (skin patch).
For instance a VDR agonist is administered, in one embodiment, intravenously in any conventional medium for intravenous injection, such as an aqueous saline medium, or in blood plasma medium. In other embodiments, administration is oral, for instance as a liquid or a pill. In other embodiments, administration is rectal, for example via a suppository containing the VDR agonist. In still other embodiments, administration is by direct infusion into a hepatic, renal, or pancreatic artery with a pharmaceutical composition that contains a vitamin D receptor agonist. In yet other embodiments, a target delivery technology is used to deliver the composition to the target tissue, for instance the liver, the kidney, or the pancreas. In one specific, non-limiting example, the vitamin D receptor agonist is designed to be taken up by the target tissue, or is linked to a target-specific carrier molecule that facilitates uptake by the target cells. For instance, for hepatic stellate cells, the vitamin D receptor agonist is conjugated to a receptor for low- and/or high-density lipoproteins (LDL and/or HDL receptors).
WO’961 at 40.
One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in formulating the agent (i.e., the VDR agonist LCA) with a pharmaceutical composition because WO’961 explains that the VDR agonist may be administered in pharmaceutical compositions. One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in formulating the pharmaceutical composition to restrict delivery of LCA to the gastrointestinal tract of the subject because WO’961 teaches rectal administration of the VDR agonist, for example, through a suppository.
Therefore, claim 20 was obvious for the reasons given above, and for the same reasons claim 1 was obvious.
Claim 21 was Obvious at the Time of Filing
Claim 21 recites the method of claim 1, wherein the diabetes is type I, type II, neonatal, or maturity onset diabetes in the young.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
The rejection set forth above is pertinent to all types of diabetes. As the Specification explains, inflammatory diseases are common complications of diabetes:
Serious long-term complications (i.e. chronic side effects) include cardiovascular disease (doubled risk), inflammatory diseases, chronic renal failure, retinal damage (which can lead to blindness), nerve damage (of several kinds), and microvascular damage, which may cause impotence and poor wound healing.
Specification at 20, paragraph[0071] (emphasis added).
WO’961 explains that hepatic fibrosis and pancreatic fibrosis are driven by inflammatory changes. See supra discussion of WO’961.
Accordingly, claim 21 was obvious for the same reasons claim 1 was obvious.
Claim 23 was Obvious at the Time of Filing
Claim 23 recites the method of claim 1, wherein the administering reduces glucose levels in the serum of a subject.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
As set forth in the rejection for claim 1, Ferrell 2019 explains that LCA is known to be a potent endogenous TGR5 agonist, and that TGR5 activation stimulates the release of GLP-1 from enteroendocrine L-cells to stimulate insulin secretion from β-cells and increase insulin sensitivity.
The Specification explains that “GLP-1 has the ability to decrease blood glucose levels in a glucose-dependent manner by enhancing insulin secretion from the pancreas.” Specification at 58, paragraph [00213].
One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in developing the method of claim 1, wherein the administering reduces glucose levels in the serum of a subject, because 1) Ferrell 2019 explains that LCA is a TGR5 agonist, and that TGR5 activation stimulates the release of GLP-1, and 2) the Specification explains that GLP-1 decreases blood glucose in a glucose-dependent manner.
Therefore, claim 23 was obvious for the reasons given above, and for the same reasons claim 1 was obvious.
Claims 24 and 25 were Obvious at the Time of Filing
Claim 24 recites the method of claim 1, wherein the subject is a mammal.
Claim 25 recites the method of claim 1, wherein the mammal is a human.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
While claim 25 is rejected as indefinite because the limitation “the mammal” lacks sufficient clarity, WO’961 explains that subjects of disclosed methods include includes both human and non-human mammals:
Subject: Living multi-cellular vertebrate organisms, a category that includes both human and non-human mammals. The methods and compositions disclosed herein have equal applications in medical and veterinary settings. Therefore, the general term "subject" is understood to include all animals, including, but not limited to, humans or veterinary subjects, such as other primates, dogs, cats, horses, and cows.
WO’961 at 15-16.
One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in developing the method of claim 1, wherein the subject is a mammal, because WO’961 explains that subjects of the disclosed methods include all animals. One having ordinary skill in the art at the time of filing would have a reasonable expectation of success in developing the method of claim 1, wherein the subject is a human, because WO’961 explains that subjects of the disclosed methods include humans.
Therefore, claims 24 and 25 were obvious for the reasons given above, and for the same reasons claim 1 was obvious.
Claim 26 was Obvious at the Time of Filing
Claim 26 recites the method of claim 1, wherein the target cell is a hepatocyte, enteroendocrine cell, an epithelial cell, an L-cell, or a neuron.
WO’961, Ferrell 2019, and Chatterjee 2005 are relied upon as above.
While claim 26 is rejected as indefinite because the limitation “the target cell” lacks sufficient clarity, WO’961 explains that hepatocytes show VDR expression:
It was previously thought that the liver lacked VDR expression because hepatocytes, the most abundant cell population in liver, usually exhibit very low levels of the receptor: the total level of VDR in rat liver is 1, 300-fold lower than in intestine.
WO’961 at 23 (emphases added).
Further, Ferrell 2019 explains that LCA is known to be a potent endogenous TGR5 agonist, and that TGR5 activation stimulates the release of GLP-1 from enteroendocrine L-cells to stimulate insulin secretion from β-cells and increase insulin sensitivity. See supra discussion of Ferrell 2019.
One having ordinary skill in the at the time of filing would have a reasonable expectation of success in developing the method of claim 1, wherein the target cell is a hepatocyte, enteroendocrine cell, an epithelial cell, an L-cell, or a neuron, because WO’961 and Ferrell 2019 explain that LCA induces activity in hepatocytes and in enteroendocrine L-cells, respectively.
Therefore, claim 26 was obvious for the reasons given above, and for the same reasons claim 1 was obvious.
Conclusion
No claims are allowed.
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/C.E.R./ Examiner, Art Unit 1629
/JEFFREY S LUNDGREN/ Supervisory Patent Examiner, Art Unit 1629
1 The examiner notes that if Applicants intend for a derivative of LCA to encompass any chemical, which the broadest reasonable interpretation of the term seems to suggest, then the claim would be rejected under 35 U.S.C. 112(d) for failing further limit the subject matter of claim 1.
2 The examiner acknowledges that the Specification provides a special definition for the term “agent”. See Specification at page 52, paragraph [00191] (“An “agent” as used herein is a chemical molecule of synthetic or biological origin.”).
3 The Specification at 20, paragraph [0071] explains that inflammatory diseases are serious long-term complications of diabetes. See Specification at 20, paragraph[0071] (“Serious long-term complications (i.e. chronic side effects) include cardiovascular disease (doubled risk), inflammatory diseases, chronic renal failure, retinal damage (which can lead to blindness), nerve damage (of several kinds), and microvascular damage, which may cause impotence and poor wound healing.”).
4 NPCs include hepatic stellate cells (HSCs), myofibroblasts, Kupffer cells (KCs), and sinusoidal endothelial cells (SECs). See WO’961 at 2.