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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 12/23/2025 has been entered.
New claims 8 and 9 are under consideration in this office action.
Withdrawn Rejection
All rejections of claim 7 are withdrawn because claim 7 has been canceled by amendment.
Claim Rejections - 35 USC § 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.
New claims 8-9 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 the following:
A method of preparing a pig model for NAFLD comprising: (a) measuring parameters in the blood of about 7 month-old micromini pig, where said blood parameters are (i) total blood cholesterol; (ii) blood albumin; (iii) HPT; (iv) blood type IV collagen; and blood hyaluronic acid; and (b) feeding the micromini pig 300/day of a CDAA diet ad libitum for a 16 weeks of time to develop symptoms of NASH ; and (c) measuring said blood parameters in the micromini pig after feeding the pigs for the 16 weeks, wherein the micromini pigs post-feeding have lipid accumulation in hepatocytes and blood profiles comprises: (1) decreased in total blood cholesterol; (2) decreased in blood albumin level; (3) decreased in HPT values; (4) increased in the blood type IV collagen level, (5) increased in blood ALT level, and (6) increased in blood hyaluronic acid level as compared to the pre-measures levels of said blood parameters, does not reasonably provide enablement for preparing a pig model for NAFLD comprising feeding any strain of pig a CDAA diet as claimed for any amount of time and developing the claims symptoms of NASH in comparison to any standard. 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 and use the invention commensurate in scope with these claims.
While determining whether a specification is enabling, one considers whether the claimed invention provides sufficient guidance to make and use the claimed invention, if not, whether an artisan would require undue experimentation to make and use the claimed invention and whether working examples have been provided. When determining whether a specification meets the enablement requirements, some of the factors that need to be analyzed are: the breadth of the claims, the nature of the invention, the state of the prior art, the level of one of ordinary skill, the level of predictability in the art, the amount of direction provided by the inventor, the existence of working examples, and whether the quantity of any necessary experimentation to make and use the invention based on the content of the disclosure is “undue”.
Nature of Invention: Producing a pig model for NAFLD by feeding a CDAA diet.
Breadth of the Claims: The claimed method encompasses feeding any strain of pig of any age any amount of a CDAA diet for any length of time, ad libitum or restricted/metered to arrive a model of NAFLD have any one of (1) decreased in total blood cholesterol; (2) decreased in blood albumin level; (3) decreased in HPT values; (4) increased in the blood type IV collagen level, (5) increased in blood ALT level, and (6) increased in blood hyaluronic acid level. The claims do not specify a reference control animal or standard by which the claimed decrease or increase is measured. As such, the breadth of the claims encompass any standard or control animal reference.
Specification Guidance: The specification generically contemplates a method preparing a NAFLD pig model that has the specific blood marker profiles of any one or more of: (1) decreased in total blood cholesterol; (2) decreased in blood albumin level; (3) decreased in HPT values; (4) increased in the blood type IV collagen level, (5) increased in blood ALT level, or (6) increased in blood hyaluronic acid level (p. 3, lines 26-35) and specifies the animal as a pig animal model (paragraph bridging page 2 and 3).
The specification states in paragraph [0020] on page 8:
In the findings (1) to (6), the “decrease” or “increase” means that the parameters corresponding to each finding is “decreased” or “increased” relative to the “standard value” in a normal individual reared using a diet that does not cause NAFLD…Examples of the diet that does not cause NAFLD, that is, a normal diet, include a solid deed MP-A…Among those skilled in the art such as sellers of experimental animals and researchers, it has been commonly recognized that there is no NAFLD finding or disease state in animals reared using a normal diet.
As such, the specification narrows the interpretation of “decrease” and “increase” in the claimed measured parameters to decreases/increases relative to a “normal individual” feed a diet that does not cause NAFLD. Therefore, the control for determining the decrease/increase in the measured parameters is in comparison to one not feed the diet. However, the breadth of the control is to any individual of any species or strain that is reared in anyway on a diet that does not cause NAFLD. The individual is referred to a “normal” however the specification does not define this term. Normative or normal is also a relative term but the specification does not define this term. As such, specification does not provide adequate guidance to determining what is “normal”. As such the standard by which the decrease/increase in paragraph [0020] on page 8 is not specifically defined and thus not specifically enabled.
Working Examples:
Example 1: Preparation of Non-Human Model Animal for NAFLD
[0035] Using micromini pigs, non-human model animals for NAFLD were prepared by the following procedure.
[0036] About 7-month-old male micromini pigs (body weight, about 10 kg) (strain: Fuji Micromini Pig) were purchased from Fuji Micra Inc., and six individuals of the pigs were acclimated to a rearing environment with natural ventilation at a temperature of 20±5° C. and a relative humidity of 55±25% under a light-dark cycle of 7:00 to 19:00 (light)/19:00 to 7:00 (dark) while being fed with 300 g/day of a solid diet MP-A (manufactured by Oriental Yeast Co., Ltd.; containing 0.29% choline and 15.3% crude protein), which is a normal diet. The feeding of the pigs was carried out at about 8:00 every day.
[0038] After an acclimation period of about 1 week, the pigs were fed ad libitum with 300 g/day of a CDAA diet (manufactured by Research Diets, Inc.; A15022101), which is substantially free of protein, in the same rearing environment for 16 weeks. The feeding of the pigs was carried out at about 8:00 every day.
Example 2: Study of Clinical Manifestations of Non-Human Model Animal for NAFLD
[0041] Micromini pigs reared as described above were studied in terms of clinical manifestations of NAFLD. As the clinical manifestations of NAFLD, the following items (1) to (6) (NAFLD findings) were set referring to indices for diagnosis of human NAFLD:
[0042] (1) a decrease in the total blood cholesterol level;
[0043] (2) a decrease in the blood albumin level;
[0044] (3) a decrease in the hepaplastin test (HPT) value;
[0045] (4) an increase in the blood type IV collagen level;
[0046] (5) an increase in the blood alanine aminotransferase (ALT) level; and
[0047] (6) an increase in the blood hyaluronic acid level.
(a) Measurement Method
[0048] During the acclimation period of about 1 week with the normal diet MP-A, which is described in Example 1, no NAFLD disease state was found. Thus, from the 6 individuals of micromini pigs after the acclimation period, blood was collected to obtain serum, and the serum samples obtained were subjected to measurement of the following items (1) to (6). The measured values were regarded as “standard values”.
[0049] (1) Total blood cholesterol level
[0050] (2) Blood albumin level
[0051] (3) Hepaplastin test (HPT) value
[0052] (4) Blood type IV collagen level
[0053] (5) Blood alanine aminotransferase (ALT) level
[0054] (6) Blood hyaluronic acid level
[0055] Each of the items (1) to (6) was measured using a method, kit, or the like described below.
[0056] (1) The total blood cholesterol level was measured by the cholesterol dehydrogenase (UV) method.
[0057] (2) The blood albumin level was measured by nephelometry (the modified BCP method).
[0058] (3) The hepaplastin test (HPT) value was measured by coagulation time measurement.
[0059] (4) The blood type IV collagen level was measured using a Type IV collagen ELISA kit, ACB (manufactured by Funakoshi Corporation).
[0060] (5) The blood alanine aminotransferase (ALT) level was measured by the JSCC reference method.
[0061] (6) The blood hyaluronic acid level was measured by the latex agglutination turbidimetric immunoassay.
[0062] Subsequently, the six individuals of micromini pigs after the acclimation period were further reared using a CDAA diet (manufactured by Research Diets, Inc.; A15022101), which is substantially free of protein. Blood was collected every 2 weeks to obtain serum during the 16 weeks after the beginning of feeding with the CDAA diet substantially free of protein. Using the serum samples obtained, the above items (1) to (6) were measured.
[0066] As can be seen from the results shown in Tables 3 and 4, all six individuals of micromini pigs reared on the CDAA diet substantially free of protein showed remarkable decreases in the blood albumin level, total cholesterol level, and hepaplastin test (HPT) value. On the other hand, they showed remarkable increases in the blood type IV collagen level, alanine aminotransferase (ALT) level, and hyaluronic acid level.
[0067] From one individual (Individual Number: 3) out of the six individuals of micromini pigs, a liver tissue was removed, and a biopsy photograph was obtained therefrom. The biopsy photograph is shown in FIG. 1.
[0068] In the biopsy photograph shown in FIG. 1, the gray areas correspond to hepatocytes, and the white small granular areas correspond to lipid (lipid droplets). The micromini-pig liver tissue obtained by the method of the present invention showed deposition of a large amount of lipid (lipid droplets) in hepatocytes, which is not found in normal liver tissues. Thus, NAFLD was found also from the biopsy photograph.
As such, the specification and particularly the working examples more narrowly provide specific guidance to a specific means of producing a NASH pig model with specially micromini pigs, under a very specific CDAA feeding protocol that results in specific blood profiles as determined by using the micropig subjects themselves as the control measure. The specification does not provide further guidance to any feeding protocol and any control as delineated in paragraph [0020] of the specification. Thus the specification does not enable the claims for the further breadth of any strain of pig, any feeding protocol and the use of any control to determine blood parameter profiles.
State of the Art: Before the time of effective filing and continuing post-filing, means of producing a NAFLD animal model was unpredictable. Flessa et al. (International Journal of Molecular Sciences 2022, 23, 15791. Pages 1-34) report, “The need for better animal models for the whole spectrum of NAFLD becomes even
more relevant for translational research, as the NAFLD prevalence is increasing worldwide due to the increasing adoption of Western-type lifestyles and eating habits. Such models would advance our understanding of the molecular mechanisms that drive NAFLD pathogenesis, as well as facilitate the ongoing research for new NAFLD/NASH treatments. HFD feeding in mice is currently a widely used approach that, in comparison to other models, closely mimics the histopathology and pathogenesis of human NAFLD whilst it also presents important features that compose the broader clinical presentation of the disease. However, HFD alone can typically lead only to hepatic steatosis, and further combined interventions are needed to proceed to NASH and HCC in the utilized animal models. Moreover, HFD feeding is a very broad term that exhibits great variations regarding the utilized diet composition and duration of feeding, while it also seems to have different effects on mice of different genetic backgrounds. Similar issues, and especially the lack of a standardized diet regimen, are raised with other dietary interventions that lead to NASH, such as the high fat, high fructose (HFHF) diet or the fast-food-like diet (High-fat/highfructose/high-cholesterol diet). Moreover, these diets and other NASH-inducing dietary models, such as the methionine and choline-deficient diet (MCD), either display discrepancies from the NASH in humans or require a long time to induce NASH features.” Page 17, 5. Conclusions, paragraph 1.
Chua et al. (International Journal of Molecules Sciences 2022, 23, 14762. Pages 1-47) report, “With high association with metabolic diseases, preclinical models for NAFLD are derived from animal models of obesity and/or insulin resistance. Over time, other types of models were developed with other methods, such as chemically induced hepatic insults and genetic models. An ideal preclinical model should meet several of these criteria: (i) closely related to the clinical etiology of the disease, (ii) closely follows the pathogenesis and features of the disease through clinical presentations, such as the presence and severity of steatosis, lobular inflammation, hepatocyte ballooning, and fibrosis, (iii) closely recapitulates the systems-level biology of the disease in humans and (iv) of suitable time cost effectiveness…. Due to the heterogeneity in NAFLD etiology, it has been a major challenge for the field to develop and identify an ideal preclinical model that could closely recapitulate the entire sequelae of NAFLD progression with strong relevance to the human phenotype…. The lack of an ideal model in this heterogeneous and complex disease resulted in NAFLD
animal model studies tending to a specific focus on a particular staging or progression of the disease and/or specific causal mechanism. Most established and widely used models have limitations in one or more features of NAFLD in humans. Although such an approach has yielded a significant understanding of disease pathology, there is still an unmet need for the successful development of therapeutics and noninvasive diagnostics. The scale of previous research performed and the lack of progress in these translational areas indicate the dire need for a closely “humanized” MAFLD animal model that is well defined.” Paragraph bridging pp. 12-13.
Regarding the production of diet-based NAFLD animals models, Chua et al. (International Journal of Molecules Sciences 2022, 23, 14762. Pages 1-47) report, “Only approximately 20% of diet-based research utilizes properly matched control
diets to their treatment counterparts…. Standard regular chow, contrary to being
“standard”, is composed of unrefined ingredients from wheat, cereal, and animal byproducts. As a control maintenance diet, it is enticing due to its widespread adoption and low cost. However, when comparing the effects of a defined MAFLD-inducing diet such as HDF with regular chow, it does little to address experimental requirements and confound experiments due to compositional inconsistencies…. Regular chow diets are vulnerable to supply chain variation, the location and conditions of harvest and processing techniques, which have a huge impact on fiber content….” Page 29, section 6.1 paragraph 1.
Thus the art before the time of filing and post-filing teaches that a predictable animal model for addressing NAFLD has not been achieved and particularly feed-based models are hindered by predictable standards and predictable controls.
As such, the breadth of the claims to a method of preparing a pig model for NAFLD with a specific phenotype of the claimed blood profile parameters by feeding any CDAA feeding protocol for any period of time in any amount measured using any control lacks enablement because the specification, particularly the working examples provides one species of making the claimed pig model with one pig strain with one feeding protocol and with the micromini pigs serving as their own control. Further, the prior art fails to supplement the shortcoming of the specification and further teaches that methods of making NAFLD animal models are unpredictably, particularly diet-induced models are hindered to predictably display the heterogenous and complex phenotype of the human disease and particularly the feed standards and controls are variable and introduce unpredictabilites in the feed-based methods of making NAFLD models.
Therefore at the time of filing the skilled artisan would need to perform an undue amount of experimentation without a predictable degree of success to implement the invention as claimed.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claim 9 is rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends. CDAA diet as claimed in independent claim 8 is recognized in the art as a specific dietary formula that is substantially protein free. As such, claim 9 does not further limit it base claim. Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
Examiner Comment
The scope indicated as enabled in the scope of enablement rejection appears to be free of the prior art.
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to MARCIA STEPHENS NOBLE whose telephone number is (571)272-5545. The examiner can normally be reached M-F 9-5:30.
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MARCIA S. NOBLE
Primary Examiner
Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632