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 1 and 3-18 are pending.
Claims 1 and 3-18, drawn to a method for prevention or treatment of a neurodegenerative disease that read on (A) SEQ ID NO: 42 as the species of peptide and (B) Parkinson as the species of neurodegenerative disease, are being acted upon in this Office Action.
Priority
Receipt is acknowledged of papers submitted under 35 U.S.C. 119(a)-(d), which papers have been placed of record in the file.
Objection Withdrawn
The objection to claims 1-14, 16-18 is withdrawn in view of the claim amendment.
The rejection of claims 2 and 8-13 under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph is withdrawn in light of the claim amendment.
Claim rejections under - 35 U.S.C. 112
The following is a quotation 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 35 U.S.C. 112 (pre-AIA ), first paragraph:
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 1 and 3-18 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 lists factors that can be used to determine if sufficient evidence of possession has been furnished in the disclosure of the application. These include "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" (MPEP 2163).
A claimed genus may be satisfied through sufficient description of a representative number of species or disclosure of relevant, identifying characteristics such as functional characteristics coupled with a known or disclosed correlation between function and structure (MPEP 2163(3)a(II)). The number of species that describe the genus must be adequate to describe the entire genus; if there is substantial variability, a large number of species must be described.
The claims encompass a method for prevention or treatment of any neurodegenerative disease of a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising: a pharmaceutically acceptable vehicle; and a therapeutically effective amount of a peptide containing comprising an amino acid sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is represented by Formula 1 below:
Formula 1
X-L-F
wherein X represents the peptide comprising an amino sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102;
L represents a linker comprising ethylene glycol repeat units;
F represents an immunoglobulin Fc region; and
"-" symbol represent a covalent linkage between X and L and between L and F, respectively.
Regarding neurodegenerative disease, the specification discloses:
[0245] As used herein, the term “neurodegenerative disease” refers to a disease that causes several symptoms resulting from degenerative changes appearing in neurons of the central nervous system and is a collective term for diseases that cause several symptoms resulting from degenerative changes appearing in neurons. Specifically, the neurodegenerative disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves.
[0246] Among the neurodegenerative diseases, degenerative brain diseases may be classified considering the main symptoms and the brain areas affected, and the degenerative brain diseases may include Parkinson’s disease (PD), Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes.
[0247] In an embodiment of the present invention, the neurodegenerative disease may be Parkinson’s disease or stroke.
[0248] In another embodiment, the neurodegenerative disease is Parkinson’s disease. Parkinson’s disease may be associated with oxidative stress, inflammatory response, apoptosis, loss of neurons, especially loss of dopaminergic neurons, e.g., loss of the substantia nigra, resulting in dopamine deficiency.
[0249] In another embodiment of the present invention, the neurodegenerative disease may be Alzheimer’s disease. “Alzheimer’s disease” is one of the most common degenerative brain diseases that cause dementia, and is a collective term for diseases that cause various symptoms resulting from degenerative changes appearing in neurons of the central nervous system. Specifically, Alzheimer’s disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Alzheimer’s disease is accompanied by cognitive function as well as neurobehavioral symptoms, such as personality changes, restlessness, depression, delusions, hallucination, increased aggression, and sleep disorders, during the progression; and neurological disorders, such as muscle rigidity and gait abnormalities, or physical complications, such as incontinence, infections, and bedsores, in the later stage. When the brain tissue of an Alzheimer’s disease patient is examined under a microscope, characteristic lesions, such as a neuritic plaque and a neurofibrillary tangle, are observed, and when observed with the naked eye, overall brain atrophy is seen due to loss of neurons. In the early stages of the disease, these brain pathological findings are mainly limited to the hippocampus, which is the main brain region responsible for memory, and the entorhinal cortex, but gradually spreads to the entire brain via the parietal lobe and frontal lobe. In accordance with the progression of the brain pathology invasion area, memory decline mainly appears in the early stage, and as it progresses, clinical symptoms vary and become more severe while showing a gradual course.
[0250] Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves. Alzheimer’s disease (AD) may also be referred to as Alzheimer’s disorder. Depending on the age of onset, Alzheimer’s disease may be divided into early-onset (elderly) Alzheimer’s disease if it develops under the age of 65, and late-onset (elderly) Alzheimer’s disease if it develops over the age of 65 years, but is not limited thereto.
[0251] Alzheimer’s disease may be associated with oxidative stress and neuronal loss.
[0252] In another embodiment, the neurodegenerative disease is progressive supranuclear palsy. Progressive supranuclear palsy may be associated with neuronal loss, particularly, loss of dopaminergic neurons.
[0253] In another embodiment, the neurodegenerative disease is multiple system atrophy. Multiple system atrophy may be associated with loss of neurons, particularly, dopaminergic neurons.
[0254] In another embodiment, the neurodegenerative disease is Lewy body dementia. Lewy body dementia may be associated with loss of neurons, particularly, dopaminergic neurons. Lewy body dementia is known to account for about 20% of the most common causes of dementia, after Alzheimer’s disease, among neurodegenerative diseases observed in the elderly. Lewy body dementia may be associated with Parkinson’s disease.
[0255] In another embodiment, the neurodegenerative disease is epilepsy. Epilepsy refers to a brain disease in which symptoms of temporary paralysis of brain functions, such as loss of consciousness, seizures, and behavioral changes, occur chronically and repeatedly, due to the temporary abnormality of brain neurons resulting in excessive excitement. In the cerebrum, neurons connected to each other exchange information through minute electrical signals. Seizures occur when these normal electrical signals are emitted abnormally and incorrectly.
[0256] In another embodiment, the neurodegenerative disease is Parkinson’s disease dementia. Parkinson’s disease may be associated with loss of neurons, particularly, dopaminergic neurons. Especially, Parkinson’s disease dementia is associated with Parkinson’s disease.
[0257] Parkinson’s disease dementia is caused by lack of dopamine due to loss of dopamine-producing cells, extensive nervous system abnormalities due to degeneration of the alpha-synuclein protein, and the like. Dopamine is an important neurotransmitter substance that acts on the basal ganglia of the brain to allow the body to move precisely as desired. Dopamine-producing cells are present in the substantia nigra in the midbrain, and when these cells are lost for a certain reason to be deficient in dopamine, resulting in movement disorders. The accumulation of Lewy bodies, generated by the denaturation of the protein called alpha-synuclein in the brain cortex causes Lewy body dementia, while the first accumulation of Lewy bodies in parts of the brain that are involved in behavior and physical functions causes Parkinson’s disease. That is, these indicate that the accumulation of the abnormal protein in the brain causes the death of brain cells, resulting in disorders in brain functions responsible for behavior and physical functions. Due to this, Parkinson’s disease patients undergo a wide range of abnormalities in the nervous system, such as hallucinations, visual hallucinations, REM sleep disorders, and olfactory disorders.
[0258] In still another embodiment, the neurodegenerative disease is stroke. Stroke may be associated with loss of neurons caused by ischemia, where ischemia may be caused by blockage (e.g., thrombosis or arterial embolism) or hemorrhage.
The specification defines prevention and treatment as follow:
[0242] As used herein, the term “prevention” refers to any action that inhibit or delay the occurrence of a neurodegenerative disease by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide, while the term “treatment” refers to any action that alleviates or advantageously change the symptoms of a neurodegenerative diseases by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide.
Regarding long-acting conjugate of Formula 1, the specification discloses:
[0150] Herein, the term “long-acting conjugate of Formula 1” refers to a form in which a peptide containing an amino acid sequence of any one of SEQ ID NOS: 1 to 102 is linked to an immunoglobulin Fc region via a linker, and the conjugate may exhibit an increase in the duration of efficacy compared with a peptide containing an amino acid sequence of any one of the amino acid sequences of SEQ ID NOS: 1 to 102, to which the immunoglobulin Fc region is not bound.
Regarding Fc region, the specification discloses:
[0193] Meanwhile, F may be an immunoglobulin Fc region and, more specifically, the immunoglobulin Fc region may be derived from IgG, but is not particularly limited thereto.
[0194] In a specific embodiment of the present invention, F (the immunoglobulin Fc region) is a dimer consisting of two polypeptide chains and has a structure in which one end of L is linked to only one polypeptide chain of the two polypeptide chains, but is not limited thereto.
[0195] In the present invention, the term “immunoglobulin Fc region” refers to a region that includes heavy chain constant region 2 (CH2) and/or heavy chain constant region 3 (CH3) portions, excluding heavy chain and light chain variable regions in an immunoglobulin. The immunoglobulin Fc region may be one element constituting a moiety of the conjugate of the present invention. The immunoglobulin Fc region may be used interchangeably with the term “immunoglobulin Fc fragment”.
[0196] Herein, the Fc region includes not only native sequences obtained by papain digestion of immunoglobulins, but also derivatives thereof, for example, variants, such as sequences in which one or more amino acid residues in the native sequence have been altered through deletion, insertion, non-conservative or conservative substitution, or a combination of these and is thus different from that of the native form. The derivatives, modifications, and variants are based on the assumption that these retain the ability to bind to FcRn. In the present invention, F may be a human immunoglobulin region, but is not limited thereto. Herein, the “biocompatible substance” or “carrier” may mean the Fc region.
[0197] F (immunoglobulin Fc region) has a structure in which two polypeptide chains are linked by a disulfide linkage, with the two chains being linked through only a nitrogen atom of one of the chains, but is not limited thereto. The linking through a nitrogen atom may be performed on the epsilon amino group or the N-terminal amino group of lysine through reductive amination.
[0223] Meanwhile, the immunoglobulin Fc region may be originated from humans, or other animals including cows, goats, pigs, mice, rabbits, hamsters, rats, and guinea pigs, and in a more specific embodiment, the immunoglobulin Fc region is originated from humans.
[0224] In addition, the immunoglobulin Fc region may be an Fc region derived from IgG, IgA, IgD, IgE, IgM, or a combination or hybrid thereof. In a still more specific embodiment, the immunoglobulin Fc region is derived from IgG or IgM, which is most abundant in the human blood, and in a still more specific embodiment, the immunoglobulin Fc region is derived from IgG, which is known to increase the half-lives of ligand-binding proteins. In a still more specific embodiment, the immunoglobulin Fc region is an IgG4 Fc region, and in a most specific embodiment, the immunoglobulin Fc region is an aglycosylated Fc region derived from human IgG4, but is not limited thereto.
[0225] In a specific embodiment, the immunoglobulin Fc region, which is a fragment of human IgG4 Fc, may be in the form of a homodimer in which two monomers are linked through a disulfide bond (inter-chain form) between cysteines, which are the third amino acids of the monomers, respectively. In particular, each monomer of the homodimer independently have/may have an inter-disulfide bond between cysteines at positions 35 and 95 and an inter-disulfide bond between cysteines at positions 141 and 199, that is, two inter-disulfide bonds (intra-chain form). With respect to the number of amino acids, each monomer may consist of 221 amino acids, and the number of the amino acids forming the homodimer may be a total of 442, but the number of amino acids is not limited thereto. Specifically, in the immunoglobulin Fc fragment, two monomers having the amino acid sequence of SEQ ID NO: 123 (consisting of 221 amino acids) form a homodimer through an inter-disulfide bond between cysteines, which are the 3rd amino acid of each monomer, wherein the monomers of the homodimer independently form an inter-disulfide bond between the cysteines at positions 35 and 95 and an inter-disulfide bond between the cysteines at positions 141 and 199, respectively, but the immunoglobulin Fc fragment is not limited thereto.
Regarding “an amino acid sequence” in claims 1, 4, 5, 6, the phrase “an amino acid sequence” encompasses a full-length sequence as well as any fragment thereof.
The specification discloses
[0303] Triple agonists showing activities for all of GLP-1, GIP, and glucagon receptors were prepared, and amino acid sequences thereof are shown in Table 1 (SEQ I D NO: 1 to 102).
However, the specification does not describe any fragment of SEQ ID NO: 102 still maintains binding to GLP-1 and GIP receptor.
Amending claims 1, 4, 5, 6 to recite “the amino acid sequence” would obviate this issue.
Example 2: Preparation of Long-Acting Conjugates of Triple Agonists
a conjugate in which a triple agonist, obtained by amidation of the C-terminus of the triple agonist of SEQ ID NO: 42, is bound to the immunoglobulin Fc via PEG was named “conjugate including SEQ ID NO: 42 and immunoglobulin Fc” or “long-acting conjugate of SEQ ID NO: 42”, and these may be used interchangeably herein.
[0322] The therapeutic effect of the triple agonist of the present invention was examined in acute and chronic animal models of Parkinson’s disease, one of the typical neurodegenerative diseases. The long-acting conjugate of the triple agonist of SEQ ID NO: 42 (long-acting conjugate of SEQ ID NO: 42) was selected as a representative example of a long-acting conjugate of a triple agonist and then experimented.
[0323] An acute Parkinson’s disease animal model was constructed by administering 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneally to C57BL/6 mice once a day for 7 days to induce the loss of dopaminergic neurons. During the experiment, the mice were housed in groups and had free access to water. Test groups were assigned into Group 1 (vehicle control group), Group 2 (MPTP-vehicle control group), and Group 3 (MPTP-long acting conjugate of SEQ ID NO: 42, 5.03 nmol/kg), with 10 mice per group. The vehicle and the long-acting conjugate of SEQ ID NO: 42 were subcutaneously administered once, 30 minutes after the first day of administration of MPTP. The experiment was terminated on the 7th day. Statistical analysis was performed by one-way ANOVA.
1) Behavioral Test (Rotarod Test)
[0324] After the experiment was completed, the symptoms of ataxia were evaluated using the Rotarod test. The mouse was allowed to move on a rotating cylinder for up to 180 seconds. The rotation speed of the cylinder was set to start at 10 rpm and gradually increase to 25 rpm after 100 seconds. After 100 seconds, the rotation speed of the cylinder was fixed at 25 rpm. The time until the mouse fell off the cylinder rotating in the above manner was measured, and a total of three trials were performed to calculate the average of the fall latency.
[0325] As a result, as can be seen in FIG. 1, the fall latency was reduced due to toxicity of administered MPTP in Group 2, while the fall latency was significantly improved in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
2) Dopamine Cell Protective Activity
[0326] After the behavioral test (Rotarod test) was completed, the mice in each group were sacrificed and brain tissue was extracted. The extracted brain tissue was immersed and fixed in a 4% paraformaldehyde solution, freshly prepared, for 24 hours, followed by hydration, and then frozen section slides were made for immunohistological staining. Subsequently, immunostaining was performed using an antibody for tyrosine hydroxylase (TH) involved in dopamine synthesis. For dopamine cell protection activity, the optical density and number of cells for stained TH in the stratum and substantia nigra were determined and quantified using Image J program.
[0327] As a result, as can be seen in FIG. 2, the number of TH-stained cells in the substantia nigra was reduced due to MPTP toxicity in Group 2, while the reduction in the number of TH-stained cells was significantly suppressed in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
[0328] A chronic Parkinson’s disease animal model was constructed by administering 25 mg/kg of MPTP and 250 mg/kg probenecid subcutaneously and intraperitoneally, respectively, to C57BL/6 mice twice a week (3.5-day interval) for a total of 5 weeks to induce the death of dopaminergic neurons. Test groups were assigned into Group 1 (vehicle control group), Group 2 (MPTP/Probenecid-vehicle control group), and Group 3 (MPTP/Probenecid-long acting conjugate of SEQ ID NO: 42, 5.03 nmol/kg), with 10 mice per group. The vehicle and the long-acting conjugate of SEQ ID NO: 42 were subcutaneously administered once a week for a total of 6 weeks after the administration of MPTP/Probenecid.
3) Effect of Inhibiting Excessive Immune Action in Parkinson’s Disease
[0329] Microglia, which are responsible for immunity in the brain, regulate immunity in the brain through phagocytosis or cytokine secretion in a situation, such as infection. Microglia in sections of the striatal region of the brain were stained using an antibody for Iba1 protein present in the microglia. The stained area was quantified using the image J program.
[0330] As a result, as can be seen in FIG. 3, the microglia were increased due to the toxicity by the administration of MPTP/Probenecid in Group 2, while the microglia were significantly reduced in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
4) Effect of Reducing Parkinson’s Disease Causative Substance
[0331] Alpha-synuclein is a representative protein that causes Parkinson’s disease. The increase and entanglement of alpha-synuclein in the brain causes the death of neurons. Among the proteins in the brain, alpha-synuclein was examined by an enzyme-linked immunoassay (ELISA) method.
[0332] As a result, as can be seen in FIG. 4, the amount of alpha-synuclein was increased due to the toxicity by the administration of MPTP/Probenecid in Group 2, while the amount of alpha-synuclein was significantly reduced in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
[0333] It could be therefore confirmed that the long-acting conjugate of SEQ ID NO: 42, a representative triple agonist of the present invention, had an effect of treating Parkinson’s disease as a representative example of neurodegenerative diseases.
Experimental Example 3: Therapeutic Effect of Long-Acting Conjugates of Triplet Agonist for Alzheimer’s Disease
[0334] In order to examine the therapeutic effect of the triple agonist of the present invention on Alzheimer’s disease, a representative neurodegenerative disease, db/db mice were used. The long-acting conjugate of the triple agonist of SEQ ID NO: 42 (long-acting conjugate of SEQ ID NO: 42) was selected as a representative example of a long-acting conjugate of a triple agonist and then experimented.
[0335] The db/db mice are used for a representative animal model of diabetes and known to show representative features of Alzheimer’s disease through may studies.
[0336] Specifically, 6-week-old db/db mice were subcutaneously administered with vehicle or the long-acting conjugate of SEQ ID NO: 42 (1.08 nmol/kg) every 2 days for a total of 12 weeks. As normal controls for the experiment, db/m mice of the same week age and db/db mice of 6 weeks of age were used. During the experiment, the mice were housed in groups and had free access to water. The test groups and control group had 7 animals for each group, and after 12-week administration, the brain tissue was extracted by autopsy and tested. Statistical analysis was performed by one-way ANOVA.
1) Amyloid Beta-Protein Reducing Effect
[0337] Amyloid beta-protein has been known to be a largest cause of Alzheimer’s disease, along with excessive phosphorylation of tau protein. After 12-week drug administration, the amount of amyloid beta 1-42 protein in the cerebral cortex of db/db mice was measured by ELISA.
[0338] As a result, as can be seen in FIG. 5, the amyloid beta 1-42 protein was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups administered vehicle.
2) Advanced Glycation End Product Reducing Effect
[0339] Advanced glycation end products (AGEs), which are sugar-modified proteins or lipids, are associated with aging and thus aggravate degenerative diseases, such as diabetes, arteriosclerosis, and Alzheimer’s disease. The advanced glycation end products in the cerebral cortex were quantified by ELISA.
[0340] As a result, as can be seen in FIG. 6, the advanced glycation end products were significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
3) Anti-Inflammatory Effect
[0341] Upon completion of 12-week administration, the anti-inflammatory effect was evaluated by measuring inflammatory cytokines in the extracted cerebral cortical tissue. Interleukin-1 beta among the inflammatory cytokines was measured by ELISA.
[0342] As a result, as can be seen in FIG. 7, the interleukin-1 beta was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
4) Protection Effect Against Oxidative Stress
[0343] 4-Hydroxynonenal (HNE), which is a byproduct produced by lipid peroxidation under oxidative stress, forms a HNE-protein conjugate by conjugation to an in vivo protein. To examine the protection effect against oxidative stress, the amount of HNE-protein conjugate in the cerebral cortex was measured by ELISA.
[0344] As a result, as can be seen in FIG. 8, the HNE-protein conjugate was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
However, there is no objective evidence of prevention or treatment of any and all neurodegenerative diseases, including but not limited to neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach, Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes or stroke by administering long-acting conjugate thereof comprising X-L-F containing peptide such as any 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102 linked to an immunoglobulin Fc region via a polyethylene glycol linker, other than treating Alzheimer’s disease or Parkinson’s disease wherein the long-acting conjugate of X-L-F wherein X is SEQ ID NO: 42, L is PEG and F is IgG1 or IgG4 Fc.
There are no objective evidence of preventing neurodegenerative diseases such as Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes or stroke.
Given the broad definition of the neurodegenerative disease and Fc (from humans, or other animals including cows, goats, pigs, mice, rabbits, hamsters, rats, and guinea pigs, variants, such as sequences in which one or more amino acid residues in the native sequence have been altered through deletion, insertion, non-conservative or conservative substitution, or a combination), the breadth of the position/location of the residues that involved one or more non-conservative or conservative substitution, deletion, addition, and a combination thereof in the Fc, the disclosure does not describe a representative number of species of such long-acting conjugates for the claimed methods.
One cannot extrapolate the teaching of the specification to the breadth of claims because it is well known that the art of preventing neurodegenerative disease is highly unpredictable, for example, Chou et al (Faculty Reviews 10(81): 1-7, 2021 PTO 892) teaches that despite progress, therapies to prevent or decrease disease progression and restore neuronal function remain a challenge and an ongoing focus in both research and clinical practice. Thus, further investigation into the neurodegenerative pathways and the identification and development of neuroprotective agents are needed to develop promising disease-modifying therapeutic approaches for the treatment of neurodegenerative disease.
In light of this, one of skilled in the art would reasonably conclude that Applicant was not in possession of the genus of long-acting conjugates for prevention or treatment of any and all neurodegenerative diseases at the time of filing. Thus, the claims lack written description.
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. Thus, the specification fails to describe these DNA sequences.
For genus claims, an adequate written description of a claimed genus requires more than a generic statement of an invention's boundaries. A patent must set forth either a representative number of species falling within the scope of the genus or structural features common to the members of the genus. Kubin, Exparte, 83 USPQ2d 1410 (Bd. Pat. App. & Int. 2007); Ariad Pharms., Inc. v. Eli Lilly& Co., 598 F.3d 1336, 1350 (Fed. Cir. 2010).
Therefore, only a method of treating Parkinson’s disease, or Alzheimer’s disease comprising administering to a subject in needed thereof an effective amount of a long-acting conjugate represent by formula I: X-L-F wherein X is a peptide consisting of the amino acid sequence of SEQ ID NO: 42, wherein L is a linker comprising polyethylene glycol of 10 kDa and F is an IgG1 Fc and a pharmaceutical acceptable vehicle, 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).
Applicants’ arguments filed May 26, 2026 have been fully considered but are not found persuasive.
Applicant has amended base claim 1 to further define the subject matter in two respects. First, the peptide X has been limited to a specific group of sequences, namely SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to 102. Second, the claim now requires that the peptide be in the form of a long-acting conjugate represented by Formula 1, X-L-F, wherein L is a linker comprising ethylene glycol repeat units and F is an immunoglobulin Fc region.
The amended claim is supported by, and consistent with, the disclosure of the specification. The representative example, SEQ ID NO: 42, is exemplified throughout the working Examples. See Specification [0309]-[0315] (preparation of long-acting conjugates of SEQ ID NOS: 21, 22, 42, 43, 50, 77, and 96); [0316]-[0321] (in vitro activities of the triple agonists and their long- acting conjugates for the GLP-1, glucagon, and GIP receptors). The remaining sequences within the recited group share substantial sequence similarity to SEQ ID NO: 42 and are characterized in the specification as triple agonists exhibiting receptor activities. See Specification [0303] and Tables 1-3.
The X-L-F structural framework is likewise expressly disclosed. See Specification [0024]-[0028] (Formula 1 and the definitions of X, L, F, and the covalent linkages). The species of L (polyethylene glycol; PEG of 1 to 100 kDa; specifically 10 kDa) and F (immunoglobulin Fc region; IgG; IgG4; aglycosylated; homodimer of two polypeptide chains, one of which is linked to L; homodimer comprising the amino acid sequence of SEQ ID NO: 123) are all described in the specification. See Specification [0035]-[0050], [0223]-[0225], [0307]. The conjugation chemistry (reaction with an amine thiol group of F and of X) is also described. See Specification [0054].
Possession of the claimed methods is further demonstrated by the working Examples, which present multiple efficacy indicators relevant to the pathology of neurodegenerative diseases using the long-acting conjugate of SEQ ID NO: 42 as a representative example. These include, in an acute and a chronic Parkinson's disease model, improvement of ataxia symptoms in the Rotarod test (Specification [0324]-[0325], FIG. 1), dopamine cell protection ( [0326]-[0327], FIG. 2), reduction of microglia ( [0329]-[0330], FIG. 3), and reduction of a-synuclein ( [0331]-[0332], FIG. 4); and in an Alzheimer's disease model, reduction of amyloid p 1-42 protein ( [0337]- [0338], FIG. 5), reduction of advanced glycation end products (AGEs) ( [0339]-[0340], FIG. 6), reduction of interleukin-13 ( [0341]-[0342], FIG. 7), and protection against oxidative stress as measured by HNE-protein conjugate ( [0343]-[0344], FIG. 8). The specification further provides the operative definitions of "prevention" and "treatment" of a neurodegenerative disease ( [0242]) and a description of the pathologies underlying the claimed indications ( [0245]-[0258]).
Taken together, the foregoing disclosure conveys to a person of ordinary skill in the art that, as of the filing date, Applicant was in possession of the methods of amended claim 1 within the entire scope now claimed, including (i) the recited group of triple-agonist peptides and (ii) the long-acting X-L-F conjugate structure for administration to a subject in need thereof.
Applicant therefore respectfully submits that the written description requirement is satisfied as to amended claim 1 and the claims that depend therefrom. Withdrawal of the rejection is respectfully requested.
In response, the amendment to claim 1 is acknowledged.
Claim 1 encompasses a method for prevention or treatment of any neurodegenerative disease of any subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising: a pharmaceutically acceptable vehicle; and a therapeutically effective amount of a peptide containing comprising an amino acid sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is represented by Formula 1 below:
Formula 1
X-L-F
wherein X represents the peptide comprising an amino sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102;
L represents a linker comprising ethylene glycol repeat units;
F represents an immunoglobulin Fc region; and
"-" symbol represent a covalent linkage between X and L and between L and F, respectively.
Applicants are not in Possession of the claimed methods for the following reasons:
Regarding “an amino acids sequence” in claims 1, 4, 5, 6, the phrase “an amino acids sequence” encompasses the full-length sequence of peptide such as any 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102 or any fragment thereof. A fragment could be as little as two amino acids.
The specification discloses
[0303] Triple agonists showing activities for all of GLP-1, GIP, and glucagon receptors were prepared, and amino acid sequences thereof are shown in Table 1 (SEQ I D NO: 1 to 102).
However, specification does not describe which fragment still maintains binding to GLP-1, CIP and glucagon receptor and function as a triple agonist for the claimed method.
Amending claims 1, 4, 5, 6 to recite “the amino acid sequence” would obviate this issue.
Regarding prevention of any neurodegenerative disease in any subject, the specification defines prevention and treatment as follow:
[0242] As used herein, the term “prevention” refers to any action that inhibit or delay the occurrence of a neurodegenerative disease by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide, while the term “treatment” refers to any action that alleviates or advantageously change the symptoms of a neurodegenerative diseases by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide.
Regarding neurodegenerative disease, the specification discloses:
[0245] As used herein, the term “neurodegenerative disease” refers to a disease that causes several symptoms resulting from degenerative changes appearing in neurons of the central nervous system and is a collective term for diseases that cause several symptoms resulting from degenerative changes appearing in neurons. Specifically, the neurodegenerative disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves.
[0246] Among the neurodegenerative diseases, degenerative brain diseases may be classified considering the main symptoms and the brain areas affected, and the degenerative brain diseases may include Parkinson’s disease (PD), Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes.
[0247] In an embodiment of the present invention, the neurodegenerative disease may be Parkinson’s disease or stroke.
[0248] In another embodiment, the neurodegenerative disease is Parkinson’s disease. Parkinson’s disease may be associated with oxidative stress, inflammatory response, apoptosis, loss of neurons, especially loss of dopaminergic neurons, e.g., loss of the substantia nigra, resulting in dopamine deficiency.
[0249] In another embodiment of the present invention, the neurodegenerative disease may be Alzheimer’s disease. “Alzheimer’s disease” is one of the most common degenerative brain diseases that cause dementia, and is a collective term for diseases that cause various symptoms resulting from degenerative changes appearing in neurons of the central nervous system. Specifically, Alzheimer’s disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Alzheimer’s disease is accompanied by cognitive function as well as neurobehavioral symptoms, such as personality changes, restlessness, depression, delusions, hallucination, increased aggression, and sleep disorders, during the progression; and neurological disorders, such as muscle rigidity and gait abnormalities, or physical complications, such as incontinence, infections, and bedsores, in the later stage. When the brain tissue of an Alzheimer’s disease patient is examined under a microscope, characteristic lesions, such as a neuritic plaque and a neurofibrillary tangle, are observed, and when observed with the naked eye, overall brain atrophy is seen due to loss of neurons. In the early stages of the disease, these brain pathological findings are mainly limited to the hippocampus, which is the main brain region responsible for memory, and the entorhinal cortex, but gradually spreads to the entire brain via the parietal lobe and frontal lobe. In accordance with the progression of the brain pathology invasion area, memory decline mainly appears in the early stage, and as it progresses, clinical symptoms vary and become more severe while showing a gradual course.
[0250] Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves. Alzheimer’s disease (AD) may also be referred to as Alzheimer’s disorder. Depending on the age of onset, Alzheimer’s disease may be divided into early-onset (elderly) Alzheimer’s disease if it develops under the age of 65, and late-onset (elderly) Alzheimer’s disease if it develops over the age of 65 years, but is not limited thereto.
Regarding subject, the specification discloses:
[0292] The subject refers to a subject suspected of having a neurodegenerative disease, and the subject suspected of having a neurodegenerative disease indicates mammals including humans, rats, livestock, and the like, which have or are at the risk of developing the corresponding disease, but any subject that can be treated with the peptide and/or conjugate or the composition containing the same is included without limitation.
However, there is no objective evidence of delaying/preventing occurrence of any and all neurodegenerative disease, including but not limited to Alzheimer's disease and Parkinson’s disease in any mammalian subject, e.g., humans, rats, livestock, and the like, but is not particularly limited thereto.
The specification discloses administering just long-acting conjugate of peptide consisting of the amino acids sequence of SEQ ID NO: 42, a PEG linker and aglycosylated Fc from IgG1 in an acute Parkinson’s disease mouse model induced by MPTP and in an Alzheimer’s disease mouse model.
The specification discloses:
[0014] FIG. 1 shows the results of behavioral (rotarod) changes of mice 7 days after administration of a long-acting conjugate of a triple conjugate in an acute Parkinson’s disease mouse model induced by MPTP (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0015] FIG. 2 shows the results obtained by measuring the inhibitory activity of the long-acting conjugate of the triple agonist on the reduction in the number of dopamine cells according to the progression of Parkinson’s disease in the substantia nigra when the brain tissue of mice was extracted 7 days after the administration of the conjugate in an acute Parkinson’s disease mouse model induced by MPTP (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0016] FIG. 3 shows the results of changes in the area occupied by microglia in the striatum, after the administration of a long-acting conjugate of a triple conjugate in a chronic Parkinson’s disease mouse model induced by MPTP/probenecid (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0017] FIG. 4 shows the results of changes in the expression level of alpha-synuclein in the striatum, after the administration of a long-acting conjugate of a triple conjugate in a chronic Parkinson’s disease mouse model induced by MPTP/probenecid (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0018] FIG. 5 shows the results of changes in amyloid beta 1-42 protein known as a causative substance of Alzheimer’s disease, after the administration of a long-acting conjugate of a triple conjugate in an Alzheimer’s disease mouse model (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0019] FIG. 6 shows the results of changes in advanced glycation end products (AGEs) in the cerebral cortex, after the administration of a long-acting conjugate of a triple conjugate in an Alzheimer’s disease mouse model (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0020] FIG. 7 confirms an anti-inflammatory effect through changes in interleukin-1 beta, an inflammatory cytokine in the cerebral cortex, after the administration of a long-acting conjugate of a triple conjugate in an Alzheimer’s disease mouse model (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
[0021] FIG. 8 confirms a protection effect against oxidative stress through changes in a 4-hydroxynoenal (HNE)-protein conjugate in the cerebral cortex, after the administration of a long-acting conjugate of a triple conjugate in an Alzheimer’s disease mouse model (*p<0.05, **p<0.01, ***p<0.001, vs. vehicle control by one-way ANOVA).
However, these two mouse models for Parkinson’s disease and Alzheimer’s disease are not representative of the genus of neurodegenerative disease as defined in the specification.
One cannot extrapolate the teaching of the specification to the breadth of claims because it is well known that the art of preventing neurodegenerative disease is highly unpredictable, for example, Chou et al (Faculty Reviews 10(81): 1-7, 2021 PTO 892) teaches that despite progress, therapies to prevent or decrease disease progression and restore neuronal function remain a challenge and an ongoing focus in both research and clinical practice. Thus, further investigation into the neurodegenerative pathways and the identification and development of neuroprotective agents are needed to develop promising disease-modifying therapeutic approaches for the treatment of neurodegenerative disease.
In light of this, one of skilled in the art would reasonably conclude that Applicant was not in possession of the genus of long-acting conjugates for prevention of any and all neurodegenerative diseases at the time of filing. Thus, the claims lack written description.
For these reasons, the rejection is maintained.
Claims 1 and 3-18 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 treating Parkinson’s disease, and Alzheimer’s disease comprising administering to a subject in needed thereof an effective amount of a long-acting conjugate represent by formula I: X-L-F wherein X is a peptide consisting of the amino acid sequence of SEQ ID NO: 42, wherein L is a linker comprising polyethylene glycol of 10 kDa and F is an Fc comprises the amino acid sequence of SEQ ID NO: 123 and a pharmaceutical acceptable vehicle, does not reasonably provide enablement for a method for “prevention” of all neurodegenerative disease, including but not limited to Parkinson’s disease, by administering any peptide such as SEQ ID NO: 42 or a long-acting conjugate thereof or treating all neurodegenerative disease, including but not limited to Parkinson’s disease, by administering peptide such as SEQ ID NO: 42. 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.
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 1 encompasses a method for prevention or treatment of any neurodegenerative disease of any subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising: a pharmaceutically acceptable vehicle; and a therapeutically effective amount of a peptide containing comprising an amino acid sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is represented by Formula 1 below:
Formula 1
X-L-F
wherein X represents the peptide comprising an amino sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102;
L represents a linker comprising ethylene glycol repeat units;
F represents an immunoglobulin Fc region; and
"-" symbol represent a covalent linkage between X and L and between L and F, respectively.
Regarding neurodegenerative disease, the specification discloses:
[0245] As used herein, the term “neurodegenerative disease” refers to a disease that causes several symptoms resulting from degenerative changes appearing in neurons of the central nervous system and is a collective term for diseases that cause several symptoms resulting from degenerative changes appearing in neurons. Specifically, the neurodegenerative disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves.
[0246] Among the neurodegenerative diseases, degenerative brain diseases may be classified considering the main symptoms and the brain areas affected, and the degenerative brain diseases may include Parkinson’s disease (PD), Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes.
[0247] In an embodiment of the present invention, the neurodegenerative disease may be Parkinson’s disease or stroke.
[0248] In another embodiment, the neurodegenerative disease is Parkinson’s disease. Parkinson’s disease may be associated with oxidative stress, inflammatory response, apoptosis, loss of neurons, especially loss of dopaminergic neurons, e.g., loss of the substantia nigra, resulting in dopamine deficiency.
[0249] In another embodiment of the present invention, the neurodegenerative disease may be Alzheimer’s disease. “Alzheimer’s disease” is one of the most common degenerative brain diseases that cause dementia, and is a collective term for diseases that cause various symptoms resulting from degenerative changes appearing in neurons of the central nervous system. Specifically, Alzheimer’s disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Alzheimer’s disease is accompanied by cognitive function as well as neurobehavioral symptoms, such as personality changes, restlessness, depression, delusions, hallucination, increased aggression, and sleep disorders, during the progression; and neurological disorders, such as muscle rigidity and gait abnormalities, or physical complications, such as incontinence, infections, and bedsores, in the later stage. When the brain tissue of an Alzheimer’s disease patient is examined under a microscope, characteristic lesions, such as a neuritic plaque and a neurofibrillary tangle, are observed, and when observed with the naked eye, overall brain atrophy is seen due to loss of neurons. In the early stages of the disease, these brain pathological findings are mainly limited to the hippocampus, which is the main brain region responsible for memory, and the entorhinal cortex, but gradually spreads to the entire brain via the parietal lobe and frontal lobe. In accordance with the progression of the brain pathology invasion area, memory decline mainly appears in the early stage, and as it progresses, clinical symptoms vary and become more severe while showing a gradual course.
[0250] Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves. Alzheimer’s disease (AD) may also be referred to as Alzheimer’s disorder. Depending on the age of onset, Alzheimer’s disease may be divided into early-onset (elderly) Alzheimer’s disease if it develops under the age of 65, and late-onset (elderly) Alzheimer’s disease if it develops over the age of 65 years, but is not limited thereto.
[0251] Alzheimer’s disease may be associated with oxidative stress and neuronal loss.
[0252] In another embodiment, the neurodegenerative disease is progressive supranuclear palsy. Progressive supranuclear palsy may be associated with neuronal loss, particularly, loss of dopaminergic neurons.
[0253] In another embodiment, the neurodegenerative disease is multiple system atrophy. Multiple system atrophy may be associated with loss of neurons, particularly, dopaminergic neurons.
[0254] In another embodiment, the neurodegenerative disease is Lewy body dementia. Lewy body dementia may be associated with loss of neurons, particularly, dopaminergic neurons. Lewy body dementia is known to account for about 20% of the most common causes of dementia, after Alzheimer’s disease, among neurodegenerative diseases observed in the elderly. Lewy body dementia may be associated with Parkinson’s disease.
[0255] In another embodiment, the neurodegenerative disease is epilepsy. Epilepsy refers to a brain disease in which symptoms of temporary paralysis of brain functions, such as loss of consciousness, seizures, and behavioral changes, occur chronically and repeatedly, due to the temporary abnormality of brain neurons resulting in excessive excitement. In the cerebrum, neurons connected to each other exchange information through minute electrical signals. Seizures occur when these normal electrical signals are emitted abnormally and incorrectly.
[0256] In another embodiment, the neurodegenerative disease is Parkinson’s disease dementia. Parkinson’s disease may be associated with loss of neurons, particularly, dopaminergic neurons. Especially, Parkinson’s disease dementia is associated with Parkinson’s disease.
[0257] Parkinson’s disease dementia is caused by lack of dopamine due to loss of dopamine-producing cells, extensive nervous system abnormalities due to degeneration of the alpha-synuclein protein, and the like. Dopamine is an important neurotransmitter substance that acts on the basal ganglia of the brain to allow the body to move precisely as desired. Dopamine-producing cells are present in the substantia nigra in the midbrain, and when these cells are lost for a certain reason to be deficient in dopamine, resulting in movement disorders. The accumulation of Lewy bodies, generated by the denaturation of the protein called alpha-synuclein in the brain cortex causes Lewy body dementia, while the first accumulation of Lewy bodies in parts of the brain that are involved in behavior and physical functions causes Parkinson’s disease. That is, these indicate that the accumulation of the abnormal protein in the brain causes the death of brain cells, resulting in disorders in brain functions responsible for behavior and physical functions. Due to this, Parkinson’s disease patients undergo a wide range of abnormalities in the nervous system, such as hallucinations, visual hallucinations, REM sleep disorders, and olfactory disorders.
[0258] In still another embodiment, the neurodegenerative disease is stroke. Stroke may be associated with loss of neurons caused by ischemia, where ischemia may be caused by blockage (e.g., thrombosis or arterial embolism) or hemorrhage.
The specification defines prevention and treatment as follow:
[0242] As used herein, the term “prevention” refers to any action that inhibit or delay the occurrence of a neurodegenerative disease by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide, while the term “treatment” refers to any action that alleviates or advantageously change the symptoms of a neurodegenerative diseases by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide.
Regarding long-acting conjugate of Formula 1, the specification discloses:
[0150] Herein, the term “long-acting conjugate of Formula 1” refers to a form in which a peptide containing an amino acid sequence of any one of SEQ ID NOS: 1 to 102 is linked to an immunoglobulin Fc region via a linker, and the conjugate may exhibit an increase in the duration of efficacy compared with a peptide containing an amino acid sequence of any one of the amino acid sequences of SEQ ID NOS: 1 to 102, to which the immunoglobulin Fc region is not bound.
Regarding Fc region, the specification discloses:
[0193] Meanwhile, F may be an immunoglobulin Fc region and, more specifically, the immunoglobulin Fc region may be derived from IgG, but is not particularly limited thereto.
[0194] In a specific embodiment of the present invention, F (the immunoglobulin Fc region) is a dimer consisting of two polypeptide chains and has a structure in which one end of L is linked to only one polypeptide chain of the two polypeptide chains, but is not limited thereto.
[0195] In the present invention, the term “immunoglobulin Fc region” refers to a region that includes heavy chain constant region 2 (CH2) and/or heavy chain constant region 3 (CH3) portions, excluding heavy chain and light chain variable regions in an immunoglobulin. The immunoglobulin Fc region may be one element constituting a moiety of the conjugate of the present invention. The immunoglobulin Fc region may be used interchangeably with the term “immunoglobulin Fc fragment”.
[0196] Herein, the Fc region includes not only native sequences obtained by papain digestion of immunoglobulins, but also derivatives thereof, for example, variants, such as sequences in which one or more amino acid residues in the native sequence have been altered through deletion, insertion, non-conservative or conservative substitution, or a combination of these and is thus different from that of the native form. The derivatives, modifications, and variants are based on the assumption that these retain the ability to bind to FcRn. In the present invention, F may be a human immunoglobulin region, but is not limited thereto. Herein, the “biocompatible substance” or “carrier” may mean the Fc region.
[0197] F (immunoglobulin Fc region) has a structure in which two polypeptide chains are linked by a disulfide linkage, with the two chains being linked through only a nitrogen atom of one of the chains, but is not limited thereto. The linking through a nitrogen atom may be performed on the epsilon amino group or the N-terminal amino group of lysine through reductive amination.
[0223] Meanwhile, the immunoglobulin Fc region may be originated from humans, or other animals including cows, goats, pigs, mice, rabbits, hamsters, rats, and guinea pigs, and in a more specific embodiment, the immunoglobulin Fc region is originated from humans.
[0224] In addition, the immunoglobulin Fc region may be an Fc region derived from IgG, IgA, IgD, IgE, IgM, or a combination or hybrid thereof. In a still more specific embodiment, the immunoglobulin Fc region is derived from IgG or IgM, which is most abundant in the human blood, and in a still more specific embodiment, the immunoglobulin Fc region is derived from IgG, which is known to increase the half-lives of ligand-binding proteins. In a still more specific embodiment, the immunoglobulin Fc region is an IgG4 Fc region, and in a most specific embodiment, the immunoglobulin Fc region is an aglycosylated Fc region derived from human IgG4, but is not limited thereto.
[0225] In a specific embodiment, the immunoglobulin Fc region, which is a fragment of human IgG4 Fc, may be in the form of a homodimer in which two monomers are linked through a disulfide bond (inter-chain form) between cysteines, which are the third amino acids of the monomers, respectively. In particular, each monomer of the homodimer independently have/may have an inter-disulfide bond between cysteines at positions 35 and 95 and an inter-disulfide bond between cysteines at positions 141 and 199, that is, two inter-disulfide bonds (intra-chain form). With respect to the number of amino acids, each monomer may consist of 221 amino acids, and the number of the amino acids forming the homodimer may be a total of 442, but the number of amino acids is not limited thereto. Specifically, in the immunoglobulin Fc fragment, two monomers having the amino acid sequence of SEQ ID NO: 123 (consisting of 221 amino acids) form a homodimer through an inter-disulfide bond between cysteines, which are the 3rd amino acid of each monomer, wherein the monomers of the homodimer independently form an inter-disulfide bond between the cysteines at positions 35 and 95 and an inter-disulfide bond between the cysteines at positions 141 and 199, respectively, but the immunoglobulin Fc fragment is not limited thereto.
The specification discloses
[0303] Triple agonists showing activities for all of GLP-1, GIP, and glucagon receptors were prepared, and amino acid sequences thereof are shown in Table 1 (SEQ I D NO: 1 to 102).
Example 2: Preparation of Long-Acting Conjugates of Triple Agonists
a conjugate in which a triple agonist, obtained by amidation of the C-terminus of the triple agonist of SEQ ID NO: 42, is bound to the immunoglobulin Fc via PEG was named “conjugate including SEQ ID NO: 42 and immunoglobulin Fc” or “long-acting conjugate of SEQ ID NO: 42”, and these may be used interchangeably herein.
[0322] The therapeutic effect of the triple agonist of the present invention was examined in acute and chronic animal models of Parkinson’s disease, one of the typical neurodegenerative diseases. The long-acting conjugate of the triple agonist of SEQ ID NO: 42 (long-acting conjugate of SEQ ID NO: 42) was selected as a representative example of a long-acting conjugate of a triple agonist and then experimented.
[0323] An acute Parkinson’s disease animal model was constructed by administering 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneally to C57BL/6 mice once a day for 7 days to induce the loss of dopaminergic neurons. During the experiment, the mice were housed in groups and had free access to water. Test groups were assigned into Group 1 (vehicle control group), Group 2 (MPTP-vehicle control group), and Group 3 (MPTP-long acting conjugate of SEQ ID NO: 42, 5.03 nmol/kg), with 10 mice per group. The vehicle and the long-acting conjugate of SEQ ID NO: 42 were subcutaneously administered once, 30 minutes after the first day of administration of MPTP. The experiment was terminated on the 7th day. Statistical analysis was performed by one-way ANOVA.
1) Behavioral Test (Rotarod Test)
[0324] After the experiment was completed, the symptoms of ataxia were evaluated using the Rotarod test. The mouse was allowed to move on a rotating cylinder for up to 180 seconds. The rotation speed of the cylinder was set to start at 10 rpm and gradually increase to 25 rpm after 100 seconds. After 100 seconds, the rotation speed of the cylinder was fixed at 25 rpm. The time until the mouse fell off the cylinder rotating in the above manner was measured, and a total of three trials were performed to calculate the average of the fall latency.
[0325] As a result, as can be seen in FIG. 1, the fall latency was reduced due to toxicity of administered MPTP in Group 2, while the fall latency was significantly improved in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
2) Dopamine Cell Protective Activity
[0326] After the behavioral test (Rotarod test) was completed, the mice in each group were sacrificed and brain tissue was extracted. The extracted brain tissue was immersed and fixed in a 4% paraformaldehyde solution, freshly prepared, for 24 hours, followed by hydration, and then frozen section slides were made for immunohistological staining. Subsequently, immunostaining was performed using an antibody for tyrosine hydroxylase (TH) involved in dopamine synthesis. For dopamine cell protection activity, the optical density and number of cells for stained TH in the stratum and substantia nigra were determined and quantified using Image J program.
[0327] As a result, as can be seen in FIG. 2, the number of TH-stained cells in the substantia nigra was reduced due to MPTP toxicity in Group 2, while the reduction in the number of TH-stained cells was significantly suppressed in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
[0328] A chronic Parkinson’s disease animal model was constructed by administering 25 mg/kg of MPTP and 250 mg/kg probenecid subcutaneously and intraperitoneally, respectively, to C57BL/6 mice twice a week (3.5-day interval) for a total of 5 weeks to induce the death of dopaminergic neurons. Test groups were assigned into Group 1 (vehicle control group), Group 2 (MPTP/Probenecid-vehicle control group), and Group 3 (MPTP/Probenecid-long acting conjugate of SEQ ID NO: 42, 5.03 nmol/kg), with 10 mice per group. The vehicle and the long-acting conjugate of SEQ ID NO: 42 were subcutaneously administered once a week for a total of 6 weeks after the administration of MPTP/Probenecid.
3) Effect of Inhibiting Excessive Immune Action in Parkinson’s Disease
[0329] Microglia, which are responsible for immunity in the brain, regulate immunity in the brain through phagocytosis or cytokine secretion in a situation, such as infection. Microglia in sections of the striatal region of the brain were stained using an antibody for Iba1 protein present in the microglia. The stained area was quantified using the image J program.
[0330] As a result, as can be seen in FIG. 3, the microglia were increased due to the toxicity by the administration of MPTP/Probenecid in Group 2, while the microglia were significantly reduced in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
4) Effect of Reducing Parkinson’s Disease Causative Substance
[0331] Alpha-synuclein is a representative protein that causes Parkinson’s disease. The increase and entanglement of alpha-synuclein in the brain causes the death of neurons. Among the proteins in the brain, alpha-synuclein was examined by an enzyme-linked immunoassay (ELISA) method.
[0332] As a result, as can be seen in FIG. 4, the amount of alpha-synuclein was increased due to the toxicity by the administration of MPTP/Probenecid in Group 2, while the amount of alpha-synuclein was significantly reduced in Group 3 receiving the long-acting conjugate of SEQ ID NO: 42.
[0333] It could be therefore confirmed that the long-acting conjugate of SEQ ID NO: 42, a representative triple agonist of the present invention, had an effect of treating Parkinson’s disease as a representative example of neurodegenerative diseases.
Experimental Example 3: Therapeutic Effect of Long-Acting Conjugates of Triplet Agonist for Alzheimer’s Disease
[0334] In order to examine the therapeutic effect of the triple agonist of the present invention on Alzheimer’s disease, a representative neurodegenerative disease, db/db mice were used. The long-acting conjugate of the triple agonist of SEQ ID NO: 42 (long-acting conjugate of SEQ ID NO: 42) was selected as a representative example of a long-acting conjugate of a triple agonist and then experimented.
[0335] The db/db mice are used for a representative animal model of diabetes and known to show representative features of Alzheimer’s disease through may studies.
[0336] Specifically, 6-week-old db/db mice were subcutaneously administered with vehicle or the long-acting conjugate of SEQ ID NO: 42 (1.08 nmol/kg) every 2 days for a total of 12 weeks. As normal controls for the experiment, db/m mice of the same week age and db/db mice of 6 weeks of age were used. During the experiment, the mice were housed in groups and had free access to water. The test groups and control group had 7 animals for each group, and after 12-week administration, the brain tissue was extracted by autopsy and tested. Statistical analysis was performed by one-way ANOVA.
1) Amyloid Beta-Protein Reducing Effect
[0337] Amyloid beta-protein has been known to be a largest cause of Alzheimer’s disease, along with excessive phosphorylation of tau protein. After 12-week drug administration, the amount of amyloid beta 1-42 protein in the cerebral cortex of db/db mice was measured by ELISA.
[0338] As a result, as can be seen in FIG. 5, the amyloid beta 1-42 protein was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups administered vehicle.
2) Advanced Glycation End Product Reducing Effect
[0339] Advanced glycation end products (AGEs), which are sugar-modified proteins or lipids, are associated with aging and thus aggravate degenerative diseases, such as diabetes, arteriosclerosis, and Alzheimer’s disease. The advanced glycation end products in the cerebral cortex were quantified by ELISA.
[0340] As a result, as can be seen in FIG. 6, the advanced glycation end products were significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
3) Anti-Inflammatory Effect
[0341] Upon completion of 12-week administration, the anti-inflammatory effect was evaluated by measuring inflammatory cytokines in the extracted cerebral cortical tissue. Interleukin-1 beta among the inflammatory cytokines was measured by ELISA.
[0342] As a result, as can be seen in FIG. 7, the interleukin-1 beta was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
4) Protection Effect Against Oxidative Stress
[0343] 4-Hydroxynonenal (HNE), which is a byproduct produced by lipid peroxidation under oxidative stress, forms a HNE-protein conjugate by conjugation to an in vivo protein. To examine the protection effect against oxidative stress, the amount of HNE-protein conjugate in the cerebral cortex was measured by ELISA.
[0344] As a result, as can be seen in FIG. 8, the HNE-protein conjugate was significantly reduced in the group administered the long-acting conjugate of SEQ ID NO: 42 compared with the control groups receiving vehicle.
However, there is no objective evidence of preventing any and all neurodegenerative diseases, including but not limited to Parkinson’s disease (PD), Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes or stroke by administering any long-acting conjugate of formula I represents by X-L-F wherein X is a peptide such as any 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102, L is a linker comprising polyethylene glycol and F represents an immunoglobulin Fc.
Regarding “an amino acids sequence” in claims 1, 4, 5, 6, the phrase “an amino acids sequence” encompasses the full-length sequence of peptide such as any 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102 or any fragment thereof. A fragment could be as little as two amino acids.
The specification does not teach such fragment still maintains binding to GLP-1, CIP and glucagon receptor and function as a triple agonist for the claimed method.
One cannot extrapolate the teaching of the specification to the breadth of claims because it is well known that the art of preventing neurodegenerative disease is highly unpredictable, for example, Chou et al (Faculty Reviews 10(81): 1-7, 2021 PTO 892) teaches that despite progress, therapies to prevent or decrease disease progression and restore neuronal function remain a challenge and an ongoing focus in both research and clinical practice. Thus, further investigation into the neurodegenerative pathways and the identification and development of neuroprotective agents are needed to develop promising disease-modifying therapeutic approaches for the treatment of neurodegenerative disease.
Given the level of unpredictability in the art, it would take undue experimentation to make and use the invention for the full scope of the claims.
Applicant’s arguments filed May 26, 2026 have been fully considered but are not found persuasive.
Applicant’s position is that [A]s discussed above, present claim 1 (i) defines the peptide X with a specific group of sequences (SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to 102) and (ii) specifies the structure of the administered active agent as a long-acting conjugate of Formula 1 in which L comprises ethylene glycol repeat units and F is an immunoglobulin Fc region. The dependent claims further specify the conjugation chemistry and the configuration of the conjugate (e.g., claims 8-13). This structurally defined conjugate composition can be prepared and used by a person of ordinary skill in the art using conventional protein/peptide conjugation and pharmaceutical formulation techniques described in the specification. See Specification [0305]- [0315] (preparation of long-acting conjugates by conjugating the C-terminus-amidated triple agonist to an immunoglobulin Fc via PEG, including the maleimide-PEG-aldehyde chemistry and the molar ratio, protein concentration, and reaction conditions); [0307] (Fc region comprising the amino acid sequence of SEQ ID NO: 123); [0316]-[0321] (in vitro activity assays for GLP- 1, glucagon, and GIP receptors using CHO cell lines).
The efficacy indicators relevant to neurodegenerative diseases recited in the claims, including those recited in claims 17 and 18, can likewise be evaluated and practiced using the experimental and evaluation methods described in the specification. See Specification [0322]- [0333] (acute and chronic Parkinson's disease models, with measurement of ataxia, dopamine cell protection, microglial reduction, and a-synuclein reduction using established assays); [0334]- [0344] (Alzheimer's disease model, with measurement of amyloid p 1-42, AGEs, interleukin-1p, and HNE-protein by ELISA). The summary at Specification [0345] demonstrates that these results support that the triple-agonist peptides or long-acting conjugates thereof can effectively treat neurodegenerative diseases.
Considering the Wands factors in view of the present claims, the breadth of the claims has been substantially reduced; the nature of the claimed method is a structurally defined long-acting conjugate having a defined Fc/PEG linker architecture; the level of skill in the art of peptide-Fc conjugation is high; the specification provides extensive direction regarding the preparation and characterization of the conjugate, the species selection for L and F, the conjugation chemistry, and the in vivo evaluation protocols; and working examples are provided in two distinct neurodegenerative disease models using the representative SEQ ID NO: 42 long-acting conjugate. In view of these considerations, Applicant respectfully submits that the quantity of experimentation necessary to practice the amended claims, including amended claim 1 and the dependent claims (including claims 14 and 16, directed to Parkinson's disease and stroke), is not undue.
In response, the amendment to claim 1 is acknowledged.
Claim 1 encompasses a method for prevention or treatment of any neurodegenerative disease of any subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising: a pharmaceutically acceptable vehicle; and a therapeutically effective amount of a peptide containing comprising an amino acid sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102, wherein the peptide is in the form of a long-acting conjugate, and the long-acting conjugate is represented by Formula 1 below:
Formula 1
X-L-F
wherein X represents the peptide comprising an amino sequence of any one of SEQ ID NOS: 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102;
L represents a linker comprising ethylene glycol repeat units;
F represents an immunoglobulin Fc region; and
"-" symbol represent a covalent linkage between X and L and between L and F, respectively.
Enablement is not commensurate in scope with the claimed methods for the following reasons:
Regarding “an amino acid sequence” in claims 1, 4, 5, 6, the phrase “an amino acid sequence” encompasses a full-length sequence as well as any fragment thereof.
The specification discloses
[0303] Triple agonists showing activities for all of GLP-1, GIP, and glucagon receptors were prepared, and amino acid sequences thereof are shown in Table 1 (SEQ I D NO: 1 to 102).
However, the specification does not describe any fragment of SEQ ID NO: 102 still maintains binding to GLP-1, glucagon, and GIP receptors.
Amending claims 1, 4, 5, 6 to recite “the amino acid sequence” would obviate this issue.
Regarding prevention of any neurodegenerative disease in any subject, the specification defines prevention and treatment as follow:
[0242] As used herein, the term “prevention” refers to any action that inhibit or delay the occurrence of a neurodegenerative disease by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide, while the term “treatment” refers to any action that alleviates or advantageously change the symptoms of a neurodegenerative diseases by administration of the above peptide (e.g., the peptide itself or a long-acting conjugate form in which a biocompatible substance is bound to the peptide) or a composition containing the peptide.
Regarding neurodegenerative disease, the specification discloses:
[0245] As used herein, the term “neurodegenerative disease” refers to a disease that causes several symptoms resulting from degenerative changes appearing in neurons of the central nervous system and is a collective term for diseases that cause several symptoms resulting from degenerative changes appearing in neurons. Specifically, the neurodegenerative disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves.
[0246] Among the neurodegenerative diseases, degenerative brain diseases may be classified considering the main symptoms and the brain areas affected, and the degenerative brain diseases may include Parkinson’s disease (PD), Alzheimer’s disease, Huntington’s disease (HD) also known as Huntington’s chorea, amyotrophic lateral sclerosis (ALS) also known as Lou Gehrig’s disease, progressive supranuclear palsy (PSP), multiple system atrophy (MSA), Lewy body dementia, Parkinson’s disease dementia, epilepsy, stroke, Huntington’s chorea, cerebral hypoxia, peripheral neuropathy, memory impairment, memory loss, forgetfulness, Pick’s disease, Creutzfeldt-Jakob disease, and nerve damage caused by complications of diabetes.
[0247] In an embodiment of the present invention, the neurodegenerative disease may be Parkinson’s disease or stroke.
[0248] In another embodiment, the neurodegenerative disease is Parkinson’s disease. Parkinson’s disease may be associated with oxidative stress, inflammatory response, apoptosis, loss of neurons, especially loss of dopaminergic neurons, e.g., loss of the substantia nigra, resulting in dopamine deficiency.
[0249] In another embodiment of the present invention, the neurodegenerative disease may be Alzheimer’s disease. “Alzheimer’s disease” is one of the most common degenerative brain diseases that cause dementia, and is a collective term for diseases that cause various symptoms resulting from degenerative changes appearing in neurons of the central nervous system. Specifically, Alzheimer’s disease may include neuropathy that causes impairments in cognitive function, learning, or memory, impairments in hand/foot sensation, or malfunctions of organs including the stomach. Alzheimer’s disease is accompanied by cognitive function as well as neurobehavioral symptoms, such as personality changes, restlessness, depression, delusions, hallucination, increased aggression, and sleep disorders, during the progression; and neurological disorders, such as muscle rigidity and gait abnormalities, or physical complications, such as incontinence, infections, and bedsores, in the later stage. When the brain tissue of an Alzheimer’s disease patient is examined under a microscope, characteristic lesions, such as a neuritic plaque and a neurofibrillary tangle, are observed, and when observed with the naked eye, overall brain atrophy is seen due to loss of neurons. In the early stages of the disease, these brain pathological findings are mainly limited to the hippocampus, which is the main brain region responsible for memory, and the entorhinal cortex, but gradually spreads to the entire brain via the parietal lobe and frontal lobe. In accordance with the progression of the brain pathology invasion area, memory decline mainly appears in the early stage, and as it progresses, clinical symptoms vary and become more severe while showing a gradual course.
[0250] Specifically, the neurodegenerative disease is known to be caused by the gradual loss of functions of a specific brain cell population in the brain and spinal cord to result in the death of brain neurons, which are most important for information transmission in the brain nervous system, problems with the formation or function of synapses that transmit information between brain neurons, or an abnormal increase or decrease in electrical activity of brain nerves. Alzheimer’s disease (AD) may also be referred to as Alzheimer’s disorder. Depending on the age of onset, Alzheimer’s disease may be divided into early-onset (elderly) Alzheimer’s disease if it develops under the age of 65, and late-onset (elderly) Alzheimer’s disease if it develops over the age of 65 years, but is not limited thereto.
Regarding subject, the specification discloses:
[0292] The subject refers to a subject suspected of having a neurodegenerative disease, and the subject suspected of having a neurodegenerative disease indicates mammals including humans, rats, livestock, and the like, which have or are at the risk of developing the corresponding disease, but any subject that can be treated with the peptide and/or conjugate or the composition containing the same is included without limitation.
However, there is no objective evidence of delaying/preventing occurrence of any and all neurodegenerative disease, including but not limited to Alzheimer's disease and Parkinson’s disease in any mammalian subject, e.g., humans, rats, livestock, and the like, but is not particularly limited thereto.
The specification discloses administering just long-acting conjugate of peptide consisting of the amino acids sequence of SEQ ID NO: 42, a PEG linker and aglycosylated Fc from IgG1 in an acute Parkinson’s disease mouse model induced by MPTP or in an Alzheimer’s disease mouse model.
However, these two mouse models for Parkinson’s disease and Alzheimer’s disease are not representative of the genus of neurodegenerative disease as defined in the specification.
Regarding “an amino acids sequence” in claims 1, 4, 5, 6, the phrase “an amino acids sequence” encompasses the full-length sequence of peptide such as any 21, 22, 42, 43, 50, 64 to 73, 75 to 77, and 96 to102 or any fragment thereof. A fragment could be as little as two amino acids.
The specification does not teach such fragment still maintains binding to GLP-1, CIP and glucagon receptor and function as a triple agonist for the claimed method.
Regarding Fc variant, Moore et al (mAbs 2(2): 181-189, 2010; PTO 892) teaches that the triple mutation Ser267Glu, His268Phe, and serine 324 threonine (Ser324Thr) in the Fc has been found to largely improve CDC at the expense of reduced ADCC and ADCP via increasing the affinity to inhibitory FcγRIIb, see entire document, Table 3, p. 186, right col.
Further, Meng et al (ACS Nano 19: 307-321, 2025; PTO 892) teaches that the blood-brain barrier (BBB) remains a major obstacle for effective for effective delivery of therapeutics to treat central nervous system (CNS) disorders. Limited BBB permeation is one of the most important factors that limit the effectivity of CNS drugs, it is not clear large long-acting conjugate where the PEG linker cannot easily cross the blood-brain barrier (BBB).
One cannot extrapolate the teaching of the specification to the breadth of claims because it is well known that the art of preventing neurodegenerative disease is highly unpredictable, for example, Chou et al (Faculty Reviews 10(81): 1-7, 2021 PTO 892) teaches that despite progress, therapies to prevent or decrease disease progression and restore neuronal function remain a challenge and an ongoing focus in both research and clinical practice. Thus, further investigation into the neurodegenerative pathways and the identification and development of neuroprotective agents are needed to develop promising disease-modifying therapeutic approaches for the treatment of neurodegenerative disease.
There are insufficient in vivo working examples.
As such, it would take undue experimentation to make and use the invention for the full scope of the claims.
For these reasons, the rejection is maintained.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103(a) which forms the basis for all obviousness rejections set forth in this Office action:
(a) A patent may not be obtained though the invention is not identically disclosed or described as set forth in section 102 of this title, if the differences between the subject matter sought to be patented and the prior art are such that the subject matter as a whole would have been obvious at the time the invention was made to a person having ordinary skill in the art to which said subject matter pertains. Patentability shall not be negatived by the manner in which the invention was made.
The factual inquiries set forth in Graham v. John Deere Co., 383 U.S. 1, 148 USPQ 459 (1966), that are applied for establishing a background for determining obviousness under 35 U.S.C. 103(a) 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 under pre-AIA 35 U.S.C. 103(a), the examiner presumes that the subject matter of the various claims was commonly owned at the time any inventions covered therein were made absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and invention dates of each claim that was not commonly owned at the time a later invention was made in order for the examiner to consider the applicability of pre-AIA 35 U.S.C. 103(c) and potential pre-AIA 35 U.S.C. 102(e), (f) or (g) prior art under pre-AIA 35 U.S.C. 103(a).
Claims 1, 3-18 are rejected under 35 U.S.C. 103 as being unpatentable over Kim et al (Neuroprotective effects of HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist in the neurodegenerative disease models, European Association for the study of diabetes (EASD) 54th Annual Meeting, Berlin, German; published Oct 1, 2018; PTO 1449) in view of Oh et al (US Patent No. 10,400,020, issued September 3, 2019; PTO 892).
Regarding claims 1, 8, 10, 11, 12, 14, 15, 16, Kim teaches a method of treating a neurodegenerative disease, e.g., Parkinson’s disease in a subject such as mice C57BL/6 or Alzheimer’s disease in a subject, e.g., db/db mice comprises administering to the subject a pharmaceutical composition comprising a long-acting conjugate such as HM15211 (aka Efocipegtutide) comprising peptide GLP-1, GIP, Glucagon triple agonist, a flexible PEG linker, and aglycosylated human IgG Fc dimer having the structure below:
PNG
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703
1200
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Greyscale
Regarding claim 17, Kim teaches that HM15211 has a dopamine cell protection effect, reducing alpha-synuclein concentration, see slide below:
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687
1205
media_image2.png
Greyscale
Restoring motor function by HM15211, see slide below:
PNG
media_image3.png
665
1193
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Greyscale
Regarding claim 18, Kim teaches that HM15211 inhibits Aβ1-42 and reducing advanced glycation end products (AGEs), see slide below:
PNG
media_image4.png
642
1189
media_image4.png
Greyscale
Kim concludes:
PNG
media_image5.png
635
1192
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Greyscale
Kim does not teach that the long-acting conjugate wherein the peptide comprises an amino acid of SEQ ID NO: 42 as per claims 1, 3, 4, 5, wherein the C-terminus of the peptide is amidated as per claim 3, wherein the peptide comprises the amino acid sequence of SEQ ID NO: 21 or 22 as per claims 5, 6, wherein the amino acids at positions 16 and 20 from the N-terminus in the sequence of the peptide form a ring as per claim 7.
However, Oh teaches various long-acting conjugate of triple agonist having activities to all of glucagon/GLP-1/GIP receptors, see entire document.
The conjugate is represented by chemical formula:
X-La-F, wherein X is a peptide having activities to a glucagon receptor, a glucagon-like peptide-1 (GLP-1) receptor and a glucose-dependent insulinotropic polypeptide (GIP) receptor, such as any one of the peptide listed in Table 1), L is a linker, e.g., maleimide-PEG-aldehyde wherein PEG is 10 kDa; and F is Fc, see col. 35, Example 2, Preparation of Long-Acting Conjugates of Triple Agonists, in particular.
Regarding claims 1, 4, 5, 6, Oh teaches examples of peptide X comprises an amino acid sequence of SEQ ID NOS: 42, see col. 17, lines 36-43, in particular.
The reference SEQ ID NO: 42 is identical to the claimed SEQ ID NO: 42, as per claim 1, 4, 5, 6, see sequence alignment below:
ALIGNMENT:
Query Match 99.5%; Score 221; Length 40;
Best Local Similarity 100.0%;
Matches 40; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 YXQGTFTSDYSKYLDEKRAKEFVQWLLDHHPSSGQPPPSC 40
||||||||||||||||||||||||||||||||||||||||
Db 1 YXQGTFTSDYSKYLDEKRAKEFVQWLLDHHPSSGQPPPSC 40
Wherein X is aminoisobutyric acid (Aib), amino acids at positions 16 and 20 form a ring as per claim 7, see col. 30, Table 1, in particular.
Regarding claim 3, Oh teaches that the peptide C-terminus may be amidated, see col. 12, line 60-64, in particular.
Regarding claim 7, Oh teaches that the amino acids at positions 16 and 20 from the N-terminus in any of the sequence of the peptide form a ring, see Table 1, col. 35, lines 32-37, in particular.
Regarding claim 8, Oh teaches the linker L (col. 9, line 37) is a non-peptide polymer, e.g., polyethylene glycol, see col. 19, lines 61-63, in particular.
Regarding claim 9, Oh teaches that the molecular weight of polyethylene glycol repeat unit in L is in a range of 1 to 100 kDa, see col. 20, line 8-12, in particular.
Regarding claim 10, Oh teaches that the F is an immunoglobulin IgG Fc region is aglycosylated, see col. 23, line 10-13, in particular.
Regarding claim 11, Oh teaches that the IgG Fc region, see col. 23, lines 3-9, in particular.
Regarding claim 12, Oh teaches that the immunoglobulin Fc region is a dimer of IgG Fc (see col. 23, line 15-20, in particular) and one end of the PEG linker is linked to one of the two Fc polypeptide chains, see col. 21, 51-58, Example 2, in particular.
Regarding claim 13, Oh teaches that the non-peptide polymer may include a reactive group which can be linked to F (e.g., an immunoglobulin Fc region) and X at both ends thereof, respectively, and specifically, a reactive group which can be linked to an amine group located at the N-terminus or lysine, or a thiol group of cysteine of X, or an amine group located at the N-terminus or lysine, or a thiol group of cysteine of F (e.g., an immunoglobulin Fc region), but the reactive group is not limited thereto. see col. 20, lines 17-34, in particular. when maleimide-PEG-aldehyde is used, the maleimide group may be linked to the SH group (aka thiol group of cysteine) of peptide X by a thioether bond and the aldehyde group may be linked to the NH2 terminus of the immunoglobulin Fc through reductive alkylation, see col. 21, line 5 to 18, in particular.
In view of the combined teachings of the references, it would have been prima facie obvious to a person of ordinary skill in the art before the effective filling date of the claimed invention to treat any neurodegenerative disease, e.g., Parkinson’s disease or Alzheimer’s disease by administering any long-acting conjugate comprising triple GLP-1/GIP/Glucagon receptor linked to Fc via a PEG linker just like Kim’s method with Oh’s long-acting conjugate comprising triple GLP-1/GIP/Glucagon receptor comprising SEQ ID NO: 42 conjugated to Fc via a PEG linker to arrive at the claimed invention with a reasonable expectation of success because Kim demonstrated the triple agonist has clear neuroprotective effects in mouse models by reducing alpha-synuclein concentration, reducing amyloid Aβ1-42 protein and reducing advanced glycation end products (AGEs) and chronic inflammation.
One of ordinary skill in the art would have been motivated to do so because Kim shows triple GLP-1, GLP-1/GIP/Glucagon receptor agonist showed promise as a potential treatment for Parkinson’s disease and Alzheimer’s disease.
One of ordinary skill in the art would have been motivated to do so because Oh teaches that the triple GLP-1, GLP-1/GIP/Glucagon receptor agonist has activities to all GLP-1, GIP, and glucagon receptors simultaneously and increase half-life for treating a target disease, see col. 2, lines 22-25, in particular.
In this case, simple substitution of a known long-acting conjugate of triple GLP-1/GIP/Glucagon receptor agonist for another would obtain predictable results, extending the half-life of the GLP-1/GIP/Glucagon receptor agonist for treating various diseases. KSR Int’l Co. v. Teleflex Inc., 550 U.S. 398, 416 (2007).
“The test of obviousness is not express suggestion of the cl aimed invention in any or all of the references but rather what the references taken collectively would suggest to those of ordinary skill in the art presumed to be familiar with them.” See In re Rosselet 146 USPQ 183, 186 (CCPA 1965).
“There is no requirement (under 35 USC 103(a)) that the prior art contain an express suggestion to combine known elements to achieve the claimed invention. Rather, the suggestion to combine may come from the prior art, as filtered through the knowledge of one skilled in the art.,” Motorola, Inc, v. Interdigital Tech. Corn., 43 USPQ2d 1481, 1489 (Fed. Cir. 1997).
Accordingly, the claimed invention as a whole was prima facie obvious to one of ordinary skill in the art before the effective filling date of the claimed invention especially in the absence of evidence to the contrary.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to PHUONG HUYNH whose telephone number is (571)272-0846. The examiner can normally be reached on 9:00 a.m. to 6:30 p.m. The examiner can also be reached on alternate alternative Friday from 9:00 a.m. to 5:30 p.m.
If attempts to reach the examiner by telephone are unsuccessful, the examiner's supervisor, Misook Yu, can be reached at 571-270-3497. The fax phone number for the organization where this application or proceeding is assigned is 571-272-0839.
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/PHUONG HUYNH/ Primary Examiner, Art Unit 1641
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