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 .
Claims 17-29 are pending and being acted upon in this Office Action.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on January 15, 2025 and December 16, 2024 have been considered by the examiner and an initialed copy of the IDS is included with this Office Action.
Specification
Applicants should amend the first line of the specification to update the relationship between the instant application and U.S. Application No. 17/742,707, filed May 12, 2022, now U.S. Patent No. 12,138,272.
The lengthy specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant's cooperation is requested in correcting any errors of which applicant may become aware in the specification.
Claim objection
Claim 28 is objected to because of the following informality: the claim uses theabbreviation ALS without first defining it. To clarify the claim, applicant should first spell out thefull term before using an abbreviation. Given the subject matter of the specification, theexaminer presumes that "ALS" stands for “Amyotrophic Lateral Sclerosis”. Appropriate correction isrequired.
Claim Rejections - 35 USC § 112
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 17-29 are 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.
The recitation of “the second dosage is lower than the first dosage” in claim 17 is indefinite and ambiguous because the “first dosage” and “second dosage” of BSEP are not defined in the claim and the specification as filed. One of ordinary skill in the art would not reasonably be apprised of the metes and bounds of the invention.
Claims 18-29 are included in the rejection because they are dependent on rejected claim and do not correct the deficiency of the claim from which they depend.
Claim rejections under - 35 U.S.C. 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 17-29 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
The MPEP § 2163 lists factors that can be used to determine if sufficient evidence of possession has been furnished in the disclosure of the Application. These include: the level of skill and knowledge in the art, partial structure, physical and/or chemical properties, functional characteristics alone or coupled with a known or disclosed correlation between structure and function, and the method of making the claimed invention. Disclosure of any combination of such identifying characteristics that distinguish the claimed invention from other materials and would lead one of skill in the art to the conclusion that the applicant was in possession of the claimed species is sufficient. See Eli Lilly, 119 F.3d at 1568, 43 USPQ2d at 1406.
For claims drawn to a genus, MPEP § 2163 states that the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus, See Eli Lilly, 119 F.3d at 1568, 43 USPQ2d at 1406, M.P.E.P. § 2163, II, A, 3, (a), (ii).
Claim 17 encompasses a method of administering Taurursodiol (TURSO) and sodium phenylbutyrate to a human subject in need thereof who has received a first dosage of an inhibitor of bile salt efflux pump (BSEP), the method comprising:(a) administering to the human subject a combination of TURSO and sodium phenylbutyrate,(b) determining or having determined a first level of serum transaminases and/or bilirubin in a first biological sample from the subject, and(c) administering to the subject a second dosage of the inhibitor of BSEP, wherein the second dosage is lower than the first dosage.
Claim 18 encompasses the method of claim 17, further comprising step (d), determining or having determined a second level of serum transaminases and/or bilirubin a second biological sample from the subject.
Claim 18 encompasses the method of claim 17, further comprising step (d), determining or having determined a second level of serum transaminases and/or bilirubin a second biological sample from the subject.
Claim 19 encompasses the method of claim 18, wherein the second level of the serum transaminases and/or bilirubin is lower than the first level.
Claim 20 encompasses the method of claim 17, wherein the biological sample is a plasma or serum sample.
Claim 21 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 to about 2 grams per day.
Claim 22 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 to about 6 grams per day.
Claim 23 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 gram once a day.
Claim 24 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 gram twice a day.
Claim 25 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 grams once a day.
Claim 26 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 grams twice a day.
Claim 27 encompasses the method of claim 17, wherein the composition is administered to the subject orally or through a feeding tube.
Claim 28 encompasses the method of claim 17, wherein the subject is diagnosed with ALS.
Claim 15 encompasses the method of claim 1, wherein the subject is suspected as having ALS.
The specification discloses:
[0102] Applicant has discovered that a combination of a bile acid (e.g. TURSO) and a phenylbutyrate compound (e.g. sodium phenylbutyrate) can be used for treating one or more symptoms of ALS. Applicant has discovered that TURSO and its metabolites, ursodeoxycholic acid and glycoursodeoxycholic acid, are inhibitors of BSEP. It was also discovered that, phenylacetic acid, a metabolite of sodium phenylbutyrate was surprisingly found to inhibit BSEP. Therefore, when inhibitors of BSEP are administered concomitantly with a composition comprising a bile acid and a phenylbutyrate compound, drug-drug interactions can result leading to an exacerbation of accumulation of conjugated bile salts in the liver, thereby leading to adverse events. Accordingly, when inhibitors of BSEP and a composition comprising, e.g. TURSO and sodium phenylbutyrate are administered concomitantly to a subject in need of both treatment, signs of drug-drug interactions can be monitored, and the dose of the inhibitors of BSEP can be adjusted accordingly. For example, when BSEP inhibitors are administered concomitantly with a composition comprising TURSO and sodium phenylbutyrate, the levels of serum transaminases and bilirubin can be elevated, indicating liver toxicity. Monitoring of serum transaminases and bilirubin, and adjusting the dosage of the inhibitors of BSEP can therefore prevent or reduce the adverse effects associated with the drug-drug interaction.
[0112] Many known drugs are BSEP inhibitors. Such drugs may, in susceptible humans, cause acquired cholestasis, which rapidly resolves after the withdrawal of the drug. Exemplary inhibitors of BSEP include but are not limited to: cyclosporine, glybenclamide, rifamycin, bosentan, troglitazone, fluvastatin, ketoconazole.
[0113] As noted above, Applicant has discovered that TURSO and its metabolites, ursodeoxycholic acid and glycoursodeoxycholic acid, are inhibitors of BSEP. It was also discovered that, phenylacetic acid, a metabolite of sodium phenylbutyrate was surprisingly found to inhibit BSEP. Concomitant usage of a BSEP inhibitor and a composition comprising a bile acid and a phenylbutyrate compound can lead to drug-drug interactions resulting in an exacerbation of accumulation of conjugated bile salts in the liver, thereby leading to adverse events, e.g., the levels of serum transaminases and bilirubin can increase resulting in toxic effects.
[0115] In some embodiments, the BSEP inhibitor is cyclosporine. Cyclosporine (also referred to as Cyclosporine A) can be used for the prophylaxis of organ rejection in allogeneic kidney, liver, and heart transplants, or to prevent bone marrow transplant rejection. Cyclosporine is used for the treatment of patients with severe active rheumatoid arthritis (RA), or severe, recalcitrant, plaque psoriasis. The ophthalmic solution of cyclosporine is indicated to increase tear production in patients suffering from keratoconjunctivitis sicca. In addition, cyclosporine is approved for the treatment of steroid dependent and steroid-resistant nephrotic syndrome due to glomerular diseases which may include minimal change nephropathy, focal and segmental glomerulosclerosis or membranous glomerulonephritis. Cyclosporine is also commonly used for the treatment of various autoimmune and inflammatory conditions such as atopic dermatitis, blistering disorders, ulcerative colitis, juvenile rheumatoid arthritis, uveitis, connective tissue diseases, as well as idiopathic thrombocytopenia purpura. The subject may have received a first dosage of cyclosporine at about 0.5 to about 15 mg/kg/day of body weight (e.g., about 0.5 to about 5 mg/kg/day, about 1 to about 4 mg/kg/day, about 2.5 mg/kg/day, or about 12 to about 15 mg/kg/day). The second dosage of cyclosporine can be less than the first dosage by about 0.1 to about 14 mg/kg/day (e.g., about 0.5 to about 2.5 mg/kg/day, about 1 to about 5 mg/kg/day).
Measuring Liver Function
[0117] Drug metabolism can have an effect on liver function. Liver function tests check the levels of certain enzymes and proteins in your blood. Levels that are higher or lower than normal can indicate liver problems. For example, testing levels of one or more of the following: alanine transaminase, aspartate transaminase, alkaline phosphatase, albumin and total protein, bilirubin, gamma-glutamyltransferase, L-lactate dehydrogenase, and prothrombin time (to measure blood clotting factors).
Serum Transaminases
[0118] Serum transaminases include alanine transaminase (ALT) and aspartate transaminase (AST). Serum transaminases (also referred to as aminotransferases) are a group of enzymes that catalyze the interconversion of amino acids and oxoacids by transfer of amino groups. ALT and AST are two of the most reliable markers of hepatocellular injury or necrosis. Their levels can be elevated in a variety of hepatic disorders.
[0119] Normal ranges of ALT and AST can vary based on sex, age, and laboratory. Individuals with elevated levels of serum transaminases can be classified as “mild” (<5 times the normal range), “moderate” (5-10 times the normal range) or “marked” (>10 times the normal range). Accordingly, in some embodiments of the methods described herein, the methods include determining a first level of ALT or AST in a sample of the subject. In some instances, the first level of ALT or AST can be higher than a normal range of ALT or AST by about 5 times, by above 5-10 times, or greater than 5 times the normal range.
[0120] Methods of measuring levels of serum transaminases are well known in the art. Typically, levels of ALT and AST are measured from a patient's blood sample and commonly tested with other liver enzymes and compounds in the blood. For example, immuno/enzyme-immnoassays or liquid chromatography/tandem mass spectrometry (LC/MS) can be used.
Bilirubin
[0121] Bilirubin (BR) is a yellowish-orange compound that occurs in the normal catabolic pathway that breaks down heme in the liver of vertebrates. Elevated levels of bilirubin are indicative of liver disease. There are three types of bilirubin: unconjugated, conjugated, and total (combination of both unconjugated and conjugated). Unconjugated (also referred to as “indirect”) bilirubin is the bilirubin created from red blood cell breakdown. It travels in the blood to the liver Conjugated (also referred to as “direct”) bilirubin is the bilirubin once it reaches the liver and undergoes a chemical change. It moves to the intestines before being removed through the urine and stool. Bilirubin tests can look at unconjugated, conjugated, and total amounts of bilirubin. For example, for adults over 18 years of age, normal total bilirubin can be up to 1.2 milligrams per deciliter (mg/dl) of blood. For those under 18 years of age, the normal level can be around 1 mg/dl. Normal results for conjugated (direct) bilirubin can be less than 0.3 mg/dl.
Example 1—Assessment of Sodium Phenylbutyrate as a Substrate and/or Inhibitor of BSEP
[0182] Experiments were conducted to assess whether sodium phenylbutyrate is a substrate and/or inhibitor of BSEP. Sodium phenylbutyrate dosed at 1 and 10 μM was not detected in uptake samples, indicating sodium phenylbutyrate was not a substrate of BSEP. Sodium phenylbutyrate (25 and 250 μM) was not an inhibitor of BSEP.
Assessment of Sodium Phenylbutyrate as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0186] The ATP-dependent uptake of sodium phenylbutyrate (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles, was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 6, below. Sodium phenylbutyrate, dosed at 1 and 10 μM, was not detected in the experimental samples, indicating no uptake had occurred. Sodium phenylbutyrate was not a substrate of BSEP.
Example 2—Assessment of Phenylacetic Acid as a Substrate and/or Inhibitor of BSEP
[0188] Experiments were conducted to assess whether phenylacetic acid is a substrate and/or inhibitor of BSEP. Adenosine triphosphate (ATP)-dependent BSEP uptake activity for phenylacetic acid at 1 and 10 μM was observed >2-fold of the adenosine monophosphate (AMP) uptake, but was not inhibited by cyclosporine A, suggesting phenylacetic acid was not a substrate of BSEP. Phenylacetic acid (750 and 7500 μM) strongly inhibited the probe substrate uptake by BSEP at 7500 μM, with an estimated IC50<7500 μM.
[0194] The ATP-dependent uptake of BSEP substrate .sup.3H-TCA in BSEP vesicles, alone or with cyclosporine A (20 μM) or phenylacetic acid (750 and 7500 μM), is presented in Table 10, below. Mean ATP-dependent uptake activity for .sup.3H-TCA was 4.69 pmol/minute/mg protein, with a signal-to-noise ratio of 9.97. The .sup.3H-TCA uptake decreased to 3.49% in the presence of BSEP inhibitor cyclosporine A. Phenylacetic acid inhibited .sup.3H-TCA uptake, with 99.6% activity remaining at 750 μM but 0.00% activity remaining at 7500 μM, for an estimated IC50<7500 μM. Phenylacetic acid was, therefore, identified as an inhibitor of BSEP.
Example 3—Assessment of Phenylacetyl-L-Glutamine as a Substrate and/or Inhibitor of BSEP
[0195] Experiments were conducted to assess whether phenylacetyl-L-glutamine is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for phenylacetyl-L-glutamine at 1 and 10 μM was not observed, suggesting phenylacetyl-L-glutamine was not a substrate of BSEP. Phenylacetyl-L-glutamine (50 and 500 μM) was not an inhibitor of BSEP.
Example 4—Assessment of Tauroursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0203] Experiments were conducted to assess whether Tauroursodeoxycholic Acid is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for tauroursodeoxycholic acid at 1 and 10 μM was >2-fold of the AMP uptake and was inhibited by cyclosporine A, identifying tauroursodeoxycholic acid as a substrate of BSEP. Tauroursodeoxycholic acid (5 and 50 μM) strongly inhibited the probe substrate uptake by BSEP, with an estimated IC50<5 μM.
Assessment of Tauroursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0208] The ATP-dependent uptake of tauroursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 17, below. Mean ATP-dependent uptake activity for tauroursodeoxycholic acid at 1 and 10 μM was 14.4 and 22.4 pmol/minute/mg protein, respectively, with a signal-to-noise ratio of 3.94- and 10.6-fold over the AMP controls, respectively. Cyclosporine A treatment resulted in ≤12.6% remaining BSEP activity. These data indicated tauroursodeoxycholic acid was actively transported and was a substrate of BSEP.
Example 5—Assessment of Ursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0210] Experiments were conducted to assess whether ursodeoxycholic acid is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake of ursodeoxycholic acid at 1 and 10 μM was <2-fold above uptake in the AMP samples, which indicated the test article was not a substrate of BSEP. Ursodeoxycholic acid (50 and 500 μM) inhibited the probe substrate uptake by BSEP, with an estimated IC50<500 μM.
Assessment of Ursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0215] The ATP-dependent uptake of ursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A
[0216] (20 μM), and the data are presented in Table 21, below. Mean ATP-dependent uptake activity for ursodeoxycholic acid could not be detected at 1 μM. The ATP-dependent uptake activity at 10 μM was 0.792 pmol/minute/mg protein from a single replicate, with a signal-to-noise ratio of only 1.26-fold over the AMP controls. These data indicated ursodeoxycholic acid was not a substrate of BSEP.
Example 6—Assessment of Glycoursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0218] Experiments were conducted to assess whether glycoursodeoxycholic acid as a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for glycoursodeoxycholic acid at 1 and 10 μM was >2-fold of the AMP uptake and was inhibited by cyclosporine A, identifying glycoursodeoxycholic acid as a substrate of BSEP. Glycoursodeoxycholic acid (10 and 100 μM) strongly inhibited the probe substrate uptake by BSEP, with an estimated IC50<10 μM.
[0223] Assessment of Glycoursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0224] The ATP-dependent uptake of glycoursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 25, below. Mean ATP-dependent uptake activity for glycoursodeoxycholic acid at 1 and 10 μM was 11.2 and 14.2 pmol/minute/mg protein, respectively. The signal-to-noise ratio could not be determined as the AMP incubations were below the limit of quantitation. No remaining BSEP activity was noted following treatment with cyclosporine A. These data indicated that glycoursodeoxycholic acid was actively transported and was a substrate of BSEP.
[0226] As shown in the examples above, sodium phenylbutyrate was not identified as a substrate of BSEP transporters. Sodium phenylbutyrate (25 and 250 μM) did not inhibit BSEP transporters. Phenylacetic acid (1 and 10 μM) was not identified as a substrate of BSEP transporters. Phenylacetic acid (750 and 7500 μM) inhibited BSEP, showing the potential for drug-drug interactions. Phenylacetyl-L-glutamine was neither a substrate (1 and 10 μM) nor inhibitor (50 and 500 μM) of BSEP transporters. Tauroursodeoxycholic acid (1 and 10 μM) was a substrate of BSEP transporters. Tauroursodeoxycholic acid (5 and 50 μM) inhibited BSEP. The BSEP inhibition interaction exhibited the potential for drug-drug interactions. Ursodeoxycholic acid (1 and 10 μM) was not a substrate of BSEP transporters. Ursodeoxycholic acid (50 and 500 μM) inhibited uptake of the probe substrate for BSEP. Glycoursodeoxycholic acid (1 and 10 μM) was a substrate of BSEP transporters. Glycoursodeoxycholic acid (10 and 100 μM) inhibited BSEP transporters, showing the potential for drug-drug interactions.
However, the specification does not describe the structure of the genus of inhibitor of BSEP administering to the subject and the amount of the first and second dosage administering to the subject. One of skill in the art does not know how to extrapolate these in vitro assessment of various phenylbutyrate, Taurourodeoxycholic acid or ursodeoxycholic acid as a substrate and inhibitor BSEP inhibitor or cyclosporine A to in vivo treatment of any one or more symptoms of Amyotrophic lateral sclerosis in all human subject.
The state of the art is such that there is a high degree of genetic and clinical heterogeneity seen in amyotrophic lateral sclerosis (ALS); animal and in vitro models replicate all ALS symptoms of human illness remains an unanswered and troublesome question.
For example, Berthod (in vivo and in Vitro Models to study Amyotrophic lateral sclerosis, in Amyotrophic Lateral Sclerosis, chapter 4, 2012; PTO 1449) teaches results obtained from in vitro models may not always be relevant because they are oversimplified compared to the in vivo human situation, see p. 29, in particular.
Currently, Xu (Translational Neurodegeneration 10(29): 1-18, 2021; PTO 1449) teaches over the past two decades, more than 50 drugs have shown efficacy in extending life expectancy in preclinical animals of ALS. Yet, there is still no cure for ALS that could reverse the progression of this disorder from a clinical perspective. Riluzole and edaravone are the only two disease-modifying approved by FDA for the treatment of ALS, see p. 2, Current Clinical Treatments, Table 1, in particular. Xu teaches although a wide variety of therapeutic agents have proven effective in ALS preclinical studies and some are undergoing clinical investigation, their efficacy is still suboptimal and far from satisfactory due to the challenges regarding safe and effective delivery routes, see p. 13, left col. Clinical trials have predominantly come to disappointing results in humans because of the following reasons. First, the inadequacy of ALS models is considered as an important reason for failed clinical trials. At the preclinical level, genetically modified rodents carrying ALS mutations remain the most widely used models. However, they do not fully recapitulate the complete patho-physiological and phenotypic spectrum present in ALS patients. Second, the lack of presymptomatic biomarkers and the delay in clinical diagnosis have significantly limited the therapeutic potential of putative disease-modifying drugs.
Further, Ciccone et al (News p. 1-5, 2024; PTO 892) discloses a phase 3 PHOENIX trial (NCT05021536) of AMX0035 (marketed as Relyvrio®, comprising Sodium Phenylbutyrate and Taurursodiol), a Randomized, Double-Blind, Placebo-Controlled, Multicenter Trial to Evaluate the Safety and Efficacy of AMX0035 Versus Placebo for 48-week Treatment of Adult Patients With Amyotrophic Lateral Sclerosis (ALS), results showed no significant difference on ALSFRS-R between AMX0035-treated and placebo-treated patients over a 48-week treatment period (P = .667). In addition, investigators observed no significant differences in a subset of participants who met the CENTAUR trial (NCT03127514) criteria, the previous phase 3 study that led to the therapy’s 2022 approval. AMX0035, a coformulation of sodium phenylbutyrate and taurursodiol, maintained its safe profile, with no new safety signals observed. Relyvrio® is no longer commercially available in the United States and Canada.
There are no in vivo working examples in the specification as filed. Accordingly, the skilled artisan would recognize that applicants were NOT in possession of the invention as broadly claimed at the time the application was filed.
Vas-Cath Inc. v. Mahurkar, 19 USPQ2d 1111, makes clear that “applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the ‘written description’ inquiry, whatever is now claimed.” (see page 1117). The specification does not “clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed.” (see Vas-Cath at page 1116).
Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it. See Fiers v. Revel, 25 USPQ2d 1601, 1606 (CAFC 1993) and Amgen Inc. v. Chugai Pharmaceutical Co. Ltd., 18 USPQ2d 1016.
One cannot describe what one has not conceived. See Fiddles v. Baird, 30 USPQ2d 1481, 1483. In Fiddles v. Baird, claims directed to mammalian FGF’s were found unpatentable due to lack of written description for the broad class. The specification provided only the bovine sequence.
Therefore, only a method of monitoring drug-drug interaction in a human subject who has received a first dosage of an inhibitor of bile salt efflux pump (BSEP), the method comprising: (a) administering to the human subject a composition comprising about 1 gram of TURSO and about 3 grams of sodium phenylbutyrate, (b) determining a first level of serum transaminases and/or bilirubin in a first biological sample from the human subject, and (c) administering to the human subject a second dosage of inhibitor of BSEP wherein the inhibitor of BSEP is cyclosporine, wherein the second dosage is lower than the first dosage if the levels of serum transaminases and bilirubin are elevated, and wherein the inhibitor of BSEP is cyclosporin and the first dosage of cyclosporine is about 0.5 to about 15 mg/kg/day, but not the full breadth of the claims meets the written description provision of 35 U.S.C. § 112, first paragraph.
Applicant is reminded that Vas-Cath makes clear that the written description provision of 35 U.S.C. § 112 is severable from its enablement provision (see page 1115).
Claims 17-29 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a method of monitoring drug-drug interaction in a human subject who has received a first dosage of an inhibitor of bile salt efflux pump (BSEP), the method comprising: (a) administering to the human subject a composition comprising about 1 gram of TURSO and about 3 grams of sodium phenylbutyrate, (b) determining a first level of serum transaminases and/or bilirubin in a first biological sample from the human subject, and (c) administering to the human subject a second dosage of inhibitor of BSEP wherein the inhibitor of BSEP is cyclosporine, wherein the second dosage is lower than the first dosage if the levels of serum transaminases and bilirubin are elevated, and wherein the inhibitor of BSEP is cyclosporin and the first dosage of cyclosporine is about 0.5 to about 15 mg/kg/day, does not reasonably provide enablement for the method as set forth in claims 17-29. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make/use the invention commensurate in scope with these claims.
Enablement is considered in view of the Wands factors (MPEP 2164.01(a)). These factors include, but are not limited to: (A) The breadth of the claims; (B) The nature of the invention; (C) The state of the prior art; (D) The level of one of ordinary skill; (E) The level of predictability in the art; (F) The amount of direction provided by the inventor; (G) The existence of working examples; and (H) The quantity of experimentation needed to make or use the invention based on the content of the disclosure. . In re Wands, 858 F.2d 731, 737, 8 USPQ2d 1400, 1404 (Fed. Cir. 1988).
Claim 17 encompasses a method of administering Taurursodiol (TURSO) and sodium phenylbutyrate to a human subject in need thereof who has received a first dosage of an inhibitor of bile salt efflux pump (BSEP), the method comprising:(a) administering to the human subject a combination of TURSO and sodium phenylbutyrate,(b) determining or having determined a first level of serum transaminases and/or bilirubin in a first biological sample from the subject, and(c) administering to the subject a second dosage of the inhibitor of BSEP, wherein the second dosage is lower than the first dosage.
Claim 18 encompasses the method of claim 17, further comprising step (d), determining or having determined a second level of serum transaminases and/or bilirubin a second biological sample from the subject.
Claim 18 encompasses the method of claim 17, further comprising step (d), determining or having determined a second level of serum transaminases and/or bilirubin a second biological sample from the subject.
Claim 19 encompasses the method of claim 18, wherein the second level of the serum transaminases and/or bilirubin is lower than the first level.
Claim 20 encompasses the method of claim 17, wherein the biological sample is a plasma or serum sample.
Claim 21 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 to about 2 grams per day.
Claim 22 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 to about 6 grams per day.
Claim 23 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 gram once a day.
Claim 24 encompasses the method of claim 17, wherein the TURSO is administered at an amount of about 1 gram twice a day.
Claim 25 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 grams once a day.
Claim 26 encompasses the method of claim 17, wherein the sodium phenylbutyrate is administered at an amount of about 3 grams twice a day.
Claim 27 encompasses the method of claim 17, wherein the composition is administered to the subject orally or through a feeding tube.
Claim 28 encompasses the method of claim 17, wherein the subject is diagnosed with ALS.
Claim 15 encompasses the method of claim 1, wherein the subject is suspected as having ALS.
The specification discloses:
[0102] Applicant has discovered that a combination of a bile acid (e.g. TURSO) and a phenylbutyrate compound (e.g. sodium phenylbutyrate) can be used for treating one or more symptoms of ALS. Applicant has discovered that TURSO and its metabolites, ursodeoxycholic acid and glycoursodeoxycholic acid, are inhibitors of BSEP. It was also discovered that, phenylacetic acid, a metabolite of sodium phenylbutyrate was surprisingly found to inhibit BSEP. Therefore, when inhibitors of BSEP are administered concomitantly with a composition comprising a bile acid and a phenylbutyrate compound, drug-drug interactions can result leading to an exacerbation of accumulation of conjugated bile salts in the liver, thereby leading to adverse events. Accordingly, when inhibitors of BSEP and a composition comprising, e.g. TURSO and sodium phenylbutyrate are administered concomitantly to a subject in need of both treatment, signs of drug-drug interactions can be monitored, and the dose of the inhibitors of BSEP can be adjusted accordingly. For example, when BSEP inhibitors are administered concomitantly with a composition comprising TURSO and sodium phenylbutyrate, the levels of serum transaminases and bilirubin can be elevated, indicating liver toxicity. Monitoring of serum transaminases and bilirubin, and adjusting the dosage of the inhibitors of BSEP can therefore prevent or reduce the adverse effects associated with the drug-drug interaction.
[0112] Many known drugs are BSEP inhibitors. Such drugs may, in susceptible humans, cause acquired cholestasis, which rapidly resolves after the withdrawal of the drug. Exemplary inhibitors of BSEP include but are not limited to: cyclosporine, glybenclamide, rifamycin, bosentan, troglitazone, fluvastatin, ketoconazole.
[0113] As noted above, Applicant has discovered that TURSO and its metabolites, ursodeoxycholic acid and glycoursodeoxycholic acid, are inhibitors of BSEP. It was also discovered that, phenylacetic acid, a metabolite of sodium phenylbutyrate was surprisingly found to inhibit BSEP. Concomitant usage of a BSEP inhibitor and a composition comprising a bile acid and a phenylbutyrate compound can lead to drug-drug interactions resulting in an exacerbation of accumulation of conjugated bile salts in the liver, thereby leading to adverse events, e.g., the levels of serum transaminases and bilirubin can increase resulting in toxic effects.
[0115] In some embodiments, the BSEP inhibitor is cyclosporine. Cyclosporine (also referred to as Cyclosporine A) can be used for the prophylaxis of organ rejection in allogeneic kidney, liver, and heart transplants, or to prevent bone marrow transplant rejection. Cyclosporine is used for the treatment of patients with severe active rheumatoid arthritis (RA), or severe, recalcitrant, plaque psoriasis. The ophthalmic solution of cyclosporine is indicated to increase tear production in patients suffering from keratoconjunctivitis sicca. In addition, cyclosporine is approved for the treatment of steroid dependent and steroid-resistant nephrotic syndrome due to glomerular diseases which may include minimal change nephropathy, focal and segmental glomerulosclerosis or membranous glomerulonephritis. Cyclosporine is also commonly used for the treatment of various autoimmune and inflammatory conditions such as atopic dermatitis, blistering disorders, ulcerative colitis, juvenile rheumatoid arthritis, uveitis, connective tissue diseases, as well as idiopathic thrombocytopenia purpura. The subject may have received a first dosage of cyclosporine at about 0.5 to about 15 mg/kg/day of body weight (e.g., about 0.5 to about 5 mg/kg/day, about 1 to about 4 mg/kg/day, about 2.5 mg/kg/day, or about 12 to about 15 mg/kg/day). The second dosage of cyclosporine can be less than the first dosage by about 0.1 to about 14 mg/kg/day (e.g., about 0.5 to about 2.5 mg/kg/day, about 1 to about 5 mg/kg/day).
Measuring Liver Function
[0117] Drug metabolism can have an effect on liver function. Liver function tests check the levels of certain enzymes and proteins in your blood. Levels that are higher or lower than normal can indicate liver problems. For example, testing levels of one or more of the following: alanine transaminase, aspartate transaminase, alkaline phosphatase, albumin and total protein, bilirubin, gamma-glutamyltransferase, L-lactate dehydrogenase, and prothrombin time (to measure blood clotting factors).
Serum Transaminases
[0118] Serum transaminases include alanine transaminase (ALT) and aspartate transaminase (AST). Serum transaminases (also referred to as aminotransferases) are a group of enzymes that catalyze the interconversion of amino acids and oxoacids by transfer of amino groups. ALT and AST are two of the most reliable markers of hepatocellular injury or necrosis. Their levels can be elevated in a variety of hepatic disorders.
[0119] Normal ranges of ALT and AST can vary based on sex, age, and laboratory. Individuals with elevated levels of serum transaminases can be classified as “mild” (<5 times the normal range), “moderate” (5-10 times the normal range) or “marked” (>10 times the normal range). Accordingly, in some embodiments of the methods described herein, the methods include determining a first level of ALT or AST in a sample of the subject. In some instances, the first level of ALT or AST can be higher than a normal range of ALT or AST by about 5 times, by above 5-10 times, or greater than 5 times the normal range.
[0120] Methods of measuring levels of serum transaminases are well known in the art. Typically, levels of ALT and AST are measured from a patient's blood sample and commonly tested with other liver enzymes and compounds in the blood. For example, immuno/enzyme-immnoassays or liquid chromatography/tandem mass spectrometry (LC/MS) can be used.
Bilirubin
[0121] Bilirubin (BR) is a yellowish-orange compound that occurs in the normal catabolic pathway that breaks down heme in the liver of vertebrates. Elevated levels of bilirubin are indicative of liver disease. There are three types of bilirubin: unconjugated, conjugated, and total (combination of both unconjugated and conjugated). Unconjugated (also referred to as “indirect”) bilirubin is the bilirubin created from red blood cell breakdown. It travels in the blood to the liver Conjugated (also referred to as “direct”) bilirubin is the bilirubin once it reaches the liver and undergoes a chemical change. It moves to the intestines before being removed through the urine and stool. Bilirubin tests can look at unconjugated, conjugated, and total amounts of bilirubin. For example, for adults over 18 years of age, normal total bilirubin can be up to 1.2 milligrams per deciliter (mg/dl) of blood. For those under 18 years of age, the normal level can be around 1 mg/dl. Normal results for conjugated (direct) bilirubin can be less than 0.3 mg/dl.
Example 1—Assessment of Sodium Phenylbutyrate as a Substrate and/or Inhibitor of BSEP
[0182] Experiments were conducted to assess whether sodium phenylbutyrate is a substrate and/or inhibitor of BSEP. Sodium phenylbutyrate dosed at 1 and 10 μM was not detected in uptake samples, indicating sodium phenylbutyrate was not a substrate of BSEP. Sodium phenylbutyrate (25 and 250 μM) was not an inhibitor of BSEP.
Assessment of Sodium Phenylbutyrate as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0186] The ATP-dependent uptake of sodium phenylbutyrate (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles, was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 6, below. Sodium phenylbutyrate, dosed at 1 and 10 μM, was not detected in the experimental samples, indicating no uptake had occurred. Sodium phenylbutyrate was not a substrate of BSEP.
Example 2—Assessment of Phenylacetic Acid as a Substrate and/or Inhibitor of BSEP
[0188] Experiments were conducted to assess whether phenylacetic acid is a substrate and/or inhibitor of BSEP. Adenosine triphosphate (ATP)-dependent BSEP uptake activity for phenylacetic acid at 1 and 10 μM was observed >2-fold of the adenosine monophosphate (AMP) uptake, but was not inhibited by cyclosporine A, suggesting phenylacetic acid was not a substrate of BSEP. Phenylacetic acid (750 and 7500 μM) strongly inhibited the probe substrate uptake by BSEP at 7500 μM, with an estimated IC50<7500 μM.
[0194] The ATP-dependent uptake of BSEP substrate .sup.3H-TCA in BSEP vesicles, alone or with cyclosporine A (20 μM) or phenylacetic acid (750 and 7500 μM), is presented in Table 10, below. Mean ATP-dependent uptake activity for .sup.3H-TCA was 4.69 pmol/minute/mg protein, with a signal-to-noise ratio of 9.97. The .sup.3H-TCA uptake decreased to 3.49% in the presence of BSEP inhibitor cyclosporine A. Phenylacetic acid inhibited .sup.3H-TCA uptake, with 99.6% activity remaining at 750 μM but 0.00% activity remaining at 7500 μM, for an estimated IC50<7500 μM. Phenylacetic acid was, therefore, identified as an inhibitor of BSEP.
Example 3—Assessment of Phenylacetyl-L-Glutamine as a Substrate and/or Inhibitor of BSEP
[0195] Experiments were conducted to assess whether phenylacetyl-L-glutamine is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for phenylacetyl-L-glutamine at 1 and 10 μM was not observed, suggesting phenylacetyl-L-glutamine was not a substrate of BSEP. Phenylacetyl-L-glutamine (50 and 500 μM) was not an inhibitor of BSEP.
Example 4—Assessment of Tauroursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0203] Experiments were conducted to assess whether Tauroursodeoxycholic Acid is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for tauroursodeoxycholic acid at 1 and 10 μM was >2-fold of the AMP uptake and was inhibited by cyclosporine A, identifying tauroursodeoxycholic acid as a substrate of BSEP. Tauroursodeoxycholic acid (5 and 50 μM) strongly inhibited the probe substrate uptake by BSEP, with an estimated IC50<5 μM.
Assessment of Tauroursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0208] The ATP-dependent uptake of tauroursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 17, below. Mean ATP-dependent uptake activity for tauroursodeoxycholic acid at 1 and 10 μM was 14.4 and 22.4 pmol/minute/mg protein, respectively, with a signal-to-noise ratio of 3.94- and 10.6-fold over the AMP controls, respectively. Cyclosporine A treatment resulted in ≤12.6% remaining BSEP activity. These data indicated tauroursodeoxycholic acid was actively transported and was a substrate of BSEP.
Example 5—Assessment of Ursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0210] Experiments were conducted to assess whether ursodeoxycholic acid is a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake of ursodeoxycholic acid at 1 and 10 μM was <2-fold above uptake in the AMP samples, which indicated the test article was not a substrate of BSEP. Ursodeoxycholic acid (50 and 500 μM) inhibited the probe substrate uptake by BSEP, with an estimated IC50<500 μM.
Assessment of Ursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0215] The ATP-dependent uptake of ursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A
[0216] (20 μM), and the data are presented in Table 21, below. Mean ATP-dependent uptake activity for ursodeoxycholic acid could not be detected at 1 μM. The ATP-dependent uptake activity at 10 μM was 0.792 pmol/minute/mg protein from a single replicate, with a signal-to-noise ratio of only 1.26-fold over the AMP controls. These data indicated ursodeoxycholic acid was not a substrate of BSEP.
Example 6—Assessment of Glycoursodeoxycholic Acid as a Substrate and/or Inhibitor of BSEP
[0218] Experiments were conducted to assess whether glycoursodeoxycholic acid as a substrate and/or inhibitor of BSEP. ATP-dependent BSEP uptake activity for glycoursodeoxycholic acid at 1 and 10 μM was >2-fold of the AMP uptake and was inhibited by cyclosporine A, identifying glycoursodeoxycholic acid as a substrate of BSEP. Glycoursodeoxycholic acid (10 and 100 μM) strongly inhibited the probe substrate uptake by BSEP, with an estimated IC50<10 μM.
[0223] Assessment of Glycoursodeoxycholic Acid as a Substrate and Inhibitor of BSEP (Using Membrane Vesicles)
[0224] The ATP-dependent uptake of glycoursodeoxycholic acid (1 and 10 μM) in BSEP-transfected Sf9 membrane vesicles was tested alone or with the BSEP inhibitor cyclosporine A (20 μM), and the data are presented in Table 25, below. Mean ATP-dependent uptake activity for glycoursodeoxycholic acid at 1 and 10 μM was 11.2 and 14.2 pmol/minute/mg protein, respectively. The signal-to-noise ratio could not be determined as the AMP incubations were below the limit of quantitation. No remaining BSEP activity was noted following treatment with cyclosporine A. These data indicated that glycoursodeoxycholic acid was actively transported and was a substrate of BSEP.
[0226] As shown in the examples above, sodium phenylbutyrate was not identified as a substrate of BSEP transporters. Sodium phenylbutyrate (25 and 250 μM) did not inhibit BSEP transporters. Phenylacetic acid (1 and 10 μM) was not identified as a substrate of BSEP transporters. Phenylacetic acid (750 and 7500 μM) inhibited BSEP, showing the potential for drug-drug interactions. Phenylacetyl-L-glutamine was neither a substrate (1 and 10 μM) nor inhibitor (50 and 500 μM) of BSEP transporters. Tauroursodeoxycholic acid (1 and 10 μM) was a substrate of BSEP transporters. Tauroursodeoxycholic acid (5 and 50 μM) inhibited BSEP. The BSEP inhibition interaction exhibited the potential for drug-drug interactions. Ursodeoxycholic acid (1 and 10 μM) was not a substrate of BSEP transporters. Ursodeoxycholic acid (50 and 500 μM) inhibited uptake of the probe substrate for BSEP. Glycoursodeoxycholic acid (1 and 10 μM) was a substrate of BSEP transporters. Glycoursodeoxycholic acid (10 and 100 μM) inhibited BSEP transporters, showing the potential for drug-drug interactions.
However, the specification does not teach the chemical structures of the genus of inhibitor of BSEP administering to the subject and the amount of the first and second dosage administering to the subject. One of skill in the art how to extrapolate these in vitro assessment of various phenylbutyrate, Taurourodeoxycholic acid or ursodeoxycholic acid as a substrate and inhibitor BSEP inhibitor or cyclosporine A to in vivo treatment of any one or more symptoms of Amyotrophic lateral sclerosis in all human subject.
The state of the art is such that there is a high degree of genetic and clinical heterogeneity seen in amyotrophic lateral sclerosis (ALS); animal and in vitro models replicate all ALS symptoms of human illness remains an unanswered and troublesome question.
For example, Berthod (in vivo and in Vitro Models to study Amyotrophic lateral sclerosis, in Amyotrophic Lateral Sclerosis, chapter 4, 2012; PTO 1449) teaches results obtained from in vitro models may not always be relevant because they are oversimplified compared to the in vivo human situation, see p. 29, in particular.
Currently, Xu (Translational Neurodegeneration 10(29): 1-18, 2021; PTO 1449) teaches over the past two decades, more than 50 drugs have shown efficacy in extending life expectancy in preclinical animals of ALS. Yet, there is still no cure for ALS that could reverse the progression of this disorder from a clinical perspective. Riluzole and edaravone are the only two disease-modifying approved by FDA for the treatment of ALS, see p. 2, Current Clinical Treatments, Table 1, in particular. Xu teaches although a wide variety of therapeutic agents have proven effective in ALS preclinical studies and some are undergoing clinical investigation, their efficacy is still suboptimal and far from satisfactory due to the challenges regarding safe and effective delivery routes, see p. 13, left col. Clinical trials have predominantly come to disappointing results in humans because of the following reasons. First, the inadequacy of ALS models is considered as an important reason for failed clinical trials. At the preclinical level, genetically modified rodents carrying ALS mutations remain the most widely used models. However, they do not fully recapitulate the complete patho-physiological and phenotypic spectrum present in ALS patients. Second, the lack of presymptomatic biomarkers and the delay in clinical diagnosis have significantly limited the therapeutic potential of putative disease-modifying drugs.
Further, Ciccone et al (News p. 1-5, 2024; PTO 892) discloses a phase 3 PHOENIX trial (NCT05021536) of AMX0035 (marketed as Relyvrio®, comprising Sodium Phenylbutyrate and Taurursodiol), a Randomized, Double-Blind, Placebo-Controlled, Multicenter Trial to Evaluate the Safety and Efficacy of AMX0035 Versus Placebo for 48-week Treatment of Adult Patients With Amyotrophic Lateral Sclerosis (ALS), results showed no significant difference on ALSFRS-R between AMX0035-treated and placebo-treated patients over a 48-week treatment period (P = .667). In addition, investigators observed no significant differences in a subset of participants who met the CENTAUR trial (NCT03127514) criteria, the previous phase 3 study that led to the therap