3Notice 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 1-30 are pending.
Claims 1-30, drawn to a method of preparing anti-transferrin receptor (TfR1) antibody-PMO conjugate that read on (A) SMCC as the linker, (B) chromatography as the particular method of isolating, (C) SEQ ID NO 34 as the variable light chain and SEQ ID NO: 30 as the variable heavy chain of the particular anti-TfR1 antibody, are being acted upon in this Office Action.
Priority
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, or 365(c) is acknowledged. Applicant has not complied with one or more conditions for receiving the benefit of an earlier filing date under 35 U.S.C. 120 as follows:
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of the first paragraph of 35 U.S.C. 112. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed application, 63/327,725 fails to provide adequate support or enablement in the manner provided by the first paragraph of 35 U.S.C. 112 for one or more claims of this application. The previously-filed application does not disclose the steps of b) purifying the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof; e) capping the anti-TfR1 antibody or the anti-TfR1 antigen binding fragment thereof using an alkylating agent in claim 1, wherein the alkylating agent comprises N-ethylmaleimide (NEM) and the linker comprises a maleimide group as per claim 7, or wherein about 2.25 to 4.75 molar equivalents of PMO-linker is used in step d) and about 7.5 to 10 molar equivalents of the alkylating agent comprising NEM is used are added in step e) as per claim 9, wherein the reduced anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof is purified by filtration or chromatography as per claim 10, wherein the chromatography comprises strong cation exchange chromatography (SCX) or the filtration comprises tangential flow filtration (TFF) as per claim 11. Therefore for the purposes of applying prior art, the effective filing date of claims 1-14, 19, 23, 27-30 is April 4, 2023, the date that the application 18/130,757 was filed. The invention of claims 15-18, 20-22, 24-26 was described in application 63/327,725. Therefore the effective filing date of claims 15-18, 20-22, 24-26 is April 5, 2022, the date that the provisional application 63/327,725 was filed.
Should applicant disagree with the examiner’s factual determination above, applicant should point to evidence that shows that the invention of claims 1-30 is in fact described in one or more of the previously-filed applications.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on April 3, 2026, December 16, 2025 and September 16, 2025 have been considered by the examiner and an initialed copy of the IDS is included with this Office Action.
Drawings
The drawings filed on September 15, 2025 are acceptable.
Specification
The amendment to the specification filed on September 16, 2025 has been entered.
The lengthy specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant's cooperation is requested in correcting any errors of which applicant may become aware in the specification.
Claim Objections
Claim 1 is objected to because of the following informality: the claim uses theabbreviation PMO without first defining it. To clarify the claim, applicant should first spell out the full term before using an abbreviation. Given the subject matter of the specification, theexaminer presumes that "PMO" stands for "phosphorodiamidate morpholino oligonucleotide". Appropriate correction is required.
Claim 8 is objected to because of the following informality: the claim uses theabbreviations SMCC, MCC, MC, MBS without first defining it. To clarify the claim, applicant should first spell out the full term before using an abbreviation. Given the subject matter of the specification, theexaminer presumes that "SMCC" stands for "Succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-amidate". Appropriate correction is required.
Claim 9 is objected to because of the following informality: the claim recites “is used are added in step e)”. Appropriate correction is required.
Claim 25 is objected to because of the following informalities: “3.5- 4.5” should have been “3.5 to 4.5”.
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 1-13, 15-27, 29 and 30 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: (1) Actual reduction to practice, (2) Disclosure of drawings or structural chemical formulas, (3) Sufficient relevant identifying characteristics (such as: i. Complete structure, ii. Partial structure, iii. Physical and/or chemical properties, iv. Functional characteristics when coupled with a known or disclosed, and correlation between function and structure), (4) Method of making the claimed invention, (5) Level of skill and knowledge in the art, and (6) Predictability in the art. “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.
MPEP § 2163 states that the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus.
An adequate written description must contain enough information about the actual makeup of the claimed products – “a precise definition, such as structure, formula, chemic name, physical properties of other properties, of species falling with the genus sufficient to distinguish the gene from other materials”, which may be present in “functional terminology when the art has established a correlation between structure and function” (Amgen page 1361).
Claim 1 encompasses a method of preparing an anti-transferrin receptor (TfR1) antibody- PMO conjugate comprising the steps of:
a) contacting an anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof with a reducing agent thereby producing a reduced anti-TfR1 antibody or a reduced anti-TfR1 antigen binding fragment thereof;
b) purifying the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof;
c) contacting a PMO with a linker thereby generating a PMO-linker;
d) contacting the PMO-linker with the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof, thereby generating an anti-TfR1 antibody-PMO conjugate or an anti-TfR1 antigen binding fragment-PMO conjugate;
e) capping the anti-TfR1 antibody or the anti-TfR1 antigen binding fragment thereof using an alkylating agent; and
f) isolating the anti-TfR1 antibody-PMO conjugate or anti-TfR1 antigen binding fragment-PMO conjugate, thereby preparing the anti-TfR1 antibody-PMO conjugate or the anti-TfR1 antigen binding fragment-PMO conjugate.
Claim 2 encompasses the method of claim 1, wherein the PMO comprises the sequence of SEQ ID NO:118.
Claim 3 encompasses the method of claim 1, wherein the reducing agent comprises tris(2- carboxyethyl)phosphine (TCEP).
Claim 4 encompasses the method of claim 3, wherein about 2 to 4 molar equivalents of TCEP are used in step a).
Claim 5 encompasses the method of claim 1, wherein about 3 molar equivalents of the linker are used in step c).
Claim 6 encompasses the method of claim 1, wherein the linker is attached at the 3' terminus of the PMO.
Claim 7 encompasses the method of claim 1, wherein the alkylating agent comprises N-ethylmaleimide (NEM) and the linker comprises a maleimide group.
Claim 8 encompasses the method of claim 1, wherein the linker comprises SMCC (elected specie), MCC, MC MBS, or bismaleimide (bismal).
Claim 9 encompasses the method of claim 1, wherein about 2.25 to 4.75 molar equivalents of PMO- linker is used in step d) and about 7.5 to 10 molar equivalents of the alkylating agent comprising NEM is used are added in step e).
Claim 10 encompasses the method of claim 1, wherein the reduced anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof is purified by filtration or chromatography.
Claim 11 encompasses the method of claim 10, wherein the chromatography comprises strong cation exchange chromatography (SCX) or the filtration comprises tangential flow filtration (TFF).
Claim 12 encompasses the method of claim 1, wherein the isolated anti-TfR1 antibody-PMO conjugate or the anti-TfR1 antigen binding fragment-PMO conjugate has an average DAR ratio of about 3.5-4.5.
Claim 13 encompasses the method of claim 1, wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 34 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 30.
Claim 14 encompasses the method of claim 1, wherein the anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 63 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 48.
Claim 15 encompasses a method of preparing an antibody-PMO conjugate or an antigen binding fragment-PMO conjugate comprising the steps of:
a) contacting any antibody or antigen binding fragment thereof with a reducing agent to generate a first mixture;
b) contacting any PMO with a linker to generate a second mixture;
c) combining the first mixture and the second mixture to generate the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate; and
d) isolating the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate, thereby preparing the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate.
Claim 16 encompasses the method of claim 15, wherein the PMO comprises the sequence of SEQ ID NO:118.
Claim 17 encompasses the method of claim 15, wherein the reducing agent comprises Tris(2- carboxyethyl)phosphine (TCEP).
Claim 18 encompasses the method of claim 17, wherein about 2 to 4 molar equivalents of TCEP are used in step a).
Claim 19 encompasses the method of claim 15, wherein about 3 molar equivalents of the linker are used in step b).
Claim 20 encompasses the method of claim 15, wherein the linker is attached at the 3' terminus of the PMO.
Claim 21 encompasses the method of claim 15, wherein the linker comprises a maleimide group.
Claim 22 encompasses the method of claim 21, wherein the linker comprises SMCC (elected species), MCC, MC, MBS, or bismaleimide (bismal).
Claim 23 encompasses the method of claim 15, wherein about 2.25 to 4.75 molar equivalents of the PMO conjugated with a linker are added in step c).
Claim 24 encompasses the method of claim 15, wherein the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate is isolated by chromatography or filtration.
Claim 25 encompasses the method of claim 15, wherein the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate has an average drug to antibody ratio (DAR) of about 3.5- 4.5.
Claim 26 encompasses the method of claim 15, wherein the antibody or antigen binding fragment thereof is an anti-transferrin receptor 1 (TfR1) antibody or antigen binding fragment thereof.
Claim 27 encompasses the method of claim 26, wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 34 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 30.
Claim 28 encompasses the method of claim 26, wherein the anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 63 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 48.
Claim 29 encompasses the method of claim 26, wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19; and wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24.
Claim 30 encompasses the method of claim 1, wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19; and wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24.
The specification exemplifies anti-transferrin receptor (TfR1) antibody conjugated to just one phosphorodiamidate morpholino oligonucleotide (PMO) comprising the nucleotide sequence of SEQ ID NO: 118 (hEx44_Ac7 26-AOC). The anti-TfR1 antibody comprises a heavy chain variable region and a light chain variable region wherein the heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 30 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 34.
Scheme 1: Synthesis and Purification of hEx44_Ac7 26-AOC
[0293] An anti-human transferrin receptor antibody was produced. The hEx44_Ac7 26 PMO was synthesized by GeneTools. Antibody (10 mg/ml) in borate buffer (25 mM sodium tetraborate, 25 mM NaCl, 1 mM Diethylene triamine pentaacetic acid, pH 8.0) was reduced by adding 4 equivalents of tris (2-carboxyethyl) phosphine (TCEP) in water and incubating at 37° C. for 4 hours. 4 (N-Maleimidomethyl) cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) was coupled to the primary amine on the 3′ end of the hEx44_Ac7_26 PMO by incubating the hEx44_Ac7_26 PMO (50 mg/ml) in DMSO with 10 equivalents of SMCC (10 mg/ml) in DMSO for one hour. Unconjugated SMCC was removed by ultrafiltration using Amicon Ultra-15 centrifugal filter units with a MWCO of 3 kDa. The hEx44_Ac7_26 PMO-SMCC was washed three times with acetate buffer (10 mM sodium acetate, pH 6.0) and used immediately. The reduced antibody was mixed with 2.25 equivalents of hEx44_Ac7_26 PMO-SMCC and incubated overnight at 4° C. The pH of the reaction mixture was then reduced to 7.5 and 8 equivalents of N-Ethylmaleimide was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines.
[0294] The reaction mixture was purified with an AKTA Explorer FPLC using HIC method-1. Dependent on the conjugate, fractions containing either conjugates with a drug to antibody ratio of one (DAR 1), two (DAR 2), three (DAR 3), four (DAR 4), five (DAR 5), six (DAR 6), seven (DAR 7), eight (DAR 8) or fractions containing conjugates with a drug to antibody ratio of 3+ (DAR 3+), 4+ (DAR 4+), 5+ (DAR 5+), 6+ (DAR 6+), 7+ (DAR 7+), (DAR 8+), or fractions containing either conjugates with an average drug to antibody ratio of one (DAR 1), two (DAR 2), three (DAR 3), four (DAR 4), five (DAR 5), six (DAR 6), seven (DAR 7), or eight (DAR 8) were combined and concentrated with Amicon Ultra-15 centrifugal filter units with a MWCO of 50 kDa. Concentrated conjugates were buffer exchanged with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units prior to analysis.
[0295] Hydrophobic interaction chromatography (HIC) method-1 [0296] Column: GE, HiScreen Butyl HP, 4.7ml [0297] Solvent A: 50 mM phosphate buffer, 0.7M Ammonium Sulfate, pH 7.0; Solvent B: 80% 50 mM phosphate buffer, 20% IPA, pH 7.0; Flow Rate: 1.0 ml/min [0298] Gradient:
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Scheme 2: Synthesis and Purification of hEx44_Ac7_26-AOC
[0299] An anti-human transferrin receptor antibody was produced. The hEx44_Ac7_26 PMO was synthesized. Antibody (20.4 mg/ml) in citrate buffer (50 mM sodium citrate, 300 mM sucrose pH 6.5) was combined with ethylenediaminetetraacetic acid (EDTA, 0.5 M, 0.591 mL) and was reduced by adding 2 equivalents of tris (2-carboxyethyl) phosphine (TCEP) in water and incubating at 37° C. for 2 hours. 4 (N-Maleimidomethyl) cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) was coupled to the primary amine on the 3′ end of the hEx44_Ac7_26 PMO by incubating the hEx44_Ac7_26 PMO (50 mg/ml) in 50 mM phosphate buffer pH 7.2 with 3 equivalents of SMCC (50 mg/ml) in DMSO for one hour. Unconjugated SMCC was removed by tangential flow filtration (TFF) with a membrane MWCO of 3 kDa with acetate buffer (10 mM sodium acetate, pH 6.0). The reduced antibody was mixed with 4.75 equivalents of hEx44_Ac7_26 PMO-SMCC and incubated 1 hour at room temperature. N-Ethylmaleimide (10 equivalents, 15 mg/ml in DMSO, 25 mg) was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines. The reaction was diluted to 1 L with endotoxin free water. Excess PMO and NEM were removed via SCX purification (GE SP/HP 16 10 resin) using SCX method-1. The combined fractions were buffer exchanged via TFF into citrate buffer (50 mM sodium citrate, 60 mM NaCl, pH 5.5) and concentrated to approximately 25 mg Ab/ml. The solution was sterile filtered with a 0.22 um membrane.
[0300] Strong cation chromatography (SCX) method-1
[0301] Column: GE HiScale 50, HiPrep SP HP, 200 ml
[0302] Solvent A: 25 mM acetate, 25 mM PB, pH 6; Solvent B: 25 mM acetate, 25 mM PB, pH 6, 0.5 mM NaCl; Flow Rate: 30 ml/min
[0303] Gradient:
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Scheme 3: Synthesis and Purification of hEx44_Ac7_26-AOC
[0304] An anti-human transferrin receptor antibody was produced in citrate buffer (50 mM sodium citrate, 300 mM sucrose pH 6.5). The hEx44_Ac7_26 PMO-SMCC was synthesized by coupling 4 (N-Maleimidomethyl) cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) to the primary amine on the 3′ end of the hEx44_Ac7 26 PMO. Ethylenediaminetetraacetic acid (EDTA, 0.5 M, 0.05 mL) was added to the antibody and the solution was thoroughly mixed. The antibody was reduced by the addition of tris (2-carboxyethyl) phosphine (TCEP, 20 mg/mL in H20, 10 equiv., 19.5 mg, 0.973 mL) and the solution was incubated at 37° C. for two hours. The reduced antibody solution was removed from the incubator and cooled to room temperature. The hEx44_Ac7_26 PMO-SMCC solution (12 equiv., 27.4 mg/ml in 10 mM acetate pH 6, 26.1 mL) was added to the reduced antibody solution and mixed thoroughly. The reaction proceeded at room temperature for one hour. n-ethylmaleimide (NEM) solution (10 equiv., 25 mg/mL in DMSO, 8.5 mg, 0.34 ml) was added to the reaction mixture and the reaction proceeded for 30 minutes at room temperature to quench unreacted cysteines. The reaction mixture was diluted to 0.2 L with endotoxin free water. Excess PMO and NEM were removed via strong cation chromatography purification SCX method-1. The combined fractions were buffer exchanged via using spin filtration into histidine buffer (20 mM histidine, 10 mM methionine, 120 mM sucrose, pH 6.0) and was quantified via a BCA assay (0.79 g, 79% yield). The solution was sterile filtered with a 0.22 um membrane. PMO44-AOC was quantified via BCA and analyzed by hydrophobic interaction chromatography (average DAR≈8), size exclusion chromatography (3.3% HMW), reducing capillary gel electrophoresis (average DAR≈7.8-8.0), and ELISA binding affinity to the human transferrin receptor (74.5 pM Kd). The affinity of hEx44_Ac7_26-AOC DAR8 AOC was equivalent to unconjugated anti-transferrin receptor antibody as shown in FIG. 7.
However, the specification does not describe the structure of any and all possible antibodies (claims 15, 17-25) or anti-transferrin receptor (TfR1) antibody (claims 1, 3-12) conjugated to any and all possible PMO (claims 1, 3-15, 17-30) with any linker encompassed by the claimed methods of preparing such conjugate for treating Duchenne Muscular Dystrophy (DMD). The specification does not describe structure-identifying information, e.g., heavy and light chain variable regions or the six CDRs about all possible antibodies and the nucleotide sequences of all PMOs. The specification does not describe a representative number of species falling within the scope of the genus or structural common to the members of the genus so the one of skill in the art can visualize or recognize the member of the genus of the actual claimed anti-transferrin receptor (TfR1) antibody-PMO conjugates (claims 15, 17-25) or antibody-PMO conjugates (claims 1, 3-12) to demonstrate possession at the time of filing.
At the time of the invention was made, it is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al. taught that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences (Lloyd et al. Protein Engineering, Design & Selection 22:159-168, 2009, PTO 892; see, e.g., Discussion).
Similarly, Edwards et al., (J Mol Biol. 334(1): 103-118, 2003; PTO 892), found that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Strop et al (Chemistry and Biology 20: 161-167, 2013; PTO 892) teach drug position can have a significant effect on linker stability and antibody pharmacokinetics. The site of conjugation on the drug and antibody can influence ADC properties differently in mice and rats, highlighting potential pitfalls of examining efficacy in mouse xenograft models and toxicity in rats or nonhuman primates, see abstract, p 166, p. 168 right col, in particular.
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen (as held in Amgen), and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen (as held in Abbvie).
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 preparing an anti-transferrin receptor (TfR1) antibody- phosphorodiamidate morpholino oligonucleotide (PMO) conjugate comprising the steps of:
a) contacting an anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof with a reducing agent thereby producing a reduced anti-TfR1 antibody or a reduced anti-TfR1 antigen binding fragment thereof;
b) purifying the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof;
c) contacting a PMO with a linker thereby generating a PMO-linker;
d) contacting the PMO-linker with the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof, thereby generating an anti-TfR1 antibody-PMO conjugate or an anti-TfR1 antigen binding fragment-PMO conjugate;
e) capping the anti-TfR1 antibody or the anti-TfR1 antigen binding fragment thereof using an alkylating agent; and
f) isolating the anti-TfR1 antibody-PMO conjugate or anti-TfR1 antigen binding fragment-PMO conjugate, thereby preparing the anti-TfR1 antibody-PMO conjugate or the anti-TfR1 antigen binding fragment-PMO conjugate,
wherein the PMO comprises the sequence of SEQ ID NO:118 and
wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 34 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 30 or
wherein the anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 63 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 or
wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 17; an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 20; and an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 19; and wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises an LCDR1 consisting of the amino acid sequence of SEQ ID NO: 22; an LCDR2 consisting of the amino acid sequence of SEQ ID NO: 23; and an LCDR3 consisting of the amino acid sequence of SEQ ID NO: 24, but not the full breadth of the claims meets the written description provision of 35 U.S.C. § 112, first paragraph.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 15 and 17-26 are rejected under 35 U.S.C. 102(a)(1) or 102(a)(2) as being anticipated by Levin et al (US20180369400, published December 27, 2018; PTO 892).
Claim 15 encompasses a method of preparing an antibody-PMO conjugate or an antigen binding fragment-PMO conjugate comprising the steps of:
a) contacting any antibody or antigen binding fragment thereof with a reducing agent to generate a first mixture;
b) contacting any PMO with a linker to generate a second mixture;
c) combining the first mixture and the second mixture to generate the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate; and
d) isolating the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate, thereby preparing the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate.
Claim 17 encompasses the method of claim 15, wherein the reducing agent comprises Tris(2- carboxyethyl)phosphine (TCEP).
Claim 18 encompasses the method of claim 17, wherein about 2 to 4 molar equivalents of TCEP are used in step a).
Claim 19 encompasses the method of claim 15, wherein about 3 molar equivalents of the linker are used in step b).
Claim 20 encompasses the method of claim 15, wherein the linker is attached at the 3' terminus of the PMO.
Claim 21 encompasses the method of claim 15, wherein the linker comprises a maleimide group.
Claim 22 encompasses the method of claim 21, wherein the linker comprises SMCC (elected species).
Claim 23 encompasses the method of claim 15, wherein about 2.25 to 4.75 molar equivalents of the PMO conjugated with a linker are added in step c).
Claim 24 encompasses the method of claim 15, wherein the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate is isolated by chromatography or filtration.
Claim 25 encompasses the method of claim 15, wherein the antibody-PMO conjugate or the antigen binding fragment-PMO conjugate has an average drug to antibody ratio (DAR) of about 3.5- 4.5.
Claim 26 encompasses the method of claim 15, wherein the antibody or antigen binding fragment thereof is an anti-transferrin receptor 1 (TfR1) antibody or antigen binding fragment thereof.
Regarding claims 15, 17, 21-22, 24, and 26, Levin teaches a method of preparing an anti-transferrin receptor (TfR1) antibody-PMO conjugate, the reference method comprises: a) contacting anti-CD71 (aka transferrin receptor (TfR1) antibody) or antigen-binding fragment thereof, e.g., F(ab’)2 ([0487]) with a reducing agent by adding 4 equivalents tris(2-carboxyethyl)phosphine (TCEP) to generate a first mixture, b) contacting PMO with a linker, e.g., 4(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) to generate a mixture PMO-SMCC, c) incubate the reduced antibody and PMO-SMCC to generate antibody-PMO conjugates and then (d) isolating the anti-CD71 mAb-PMO by hydrophobic interaction chromatography (HIC), see para. [0477] to [0480].
Regarding claim 18, Levin teaches 4 molar equivalents of reducing agent, tris(2-carboxyethyl)phosphine (TCEP) was used in step a, see para. [0480].
Regarding claim 20, Levin teaches that the SMCC was coupled to the primary amine on the 3′ end of the phosphorodiamidate morpholino oligomer (PMO), see para. [0480].
Regarding claim 25, Levin teaches that the drug-to-antibody ratio is about 3, 4, 5 or greater, see para. [0342] to [0344].
Thus, the reference teachings anticipate the claimed invention.
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 15 and 27-29 are rejected under 35 U.S.C. 103 as being unpatentable over Levin et al (US20180369400, published December 27, 2018; PTO 892) in view of US patent No. 10,913,800 (issued February 9, 2021; PTO 892).
The teachings of Levin have been discussed supra.
The reference does not teach the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 34 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 30 as per claim 27 or wherein the anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 63 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 as per claim 28 or wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19; and wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24 as per claim 29.
However, the ‘800 patent teaches anti-TfR1 antibody or antigen comprising and a light variable heavy chain comprising the amino acid sequence of SEQ ID NO: 18, which is 100% identical to the claimed SEQ ID NO: 34, see col. 14, Table 14, reference SEQ ID NO: 18, 3 CDRs are underline, sequence alignment below:
Query Match 100.0%; Score 568; Length 107;
Best Local Similarity 100.0%;
Matches 107; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
Qy 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIK 107
|||||||||||||||||||||||||||||||||||||||||||||||
Db 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIK 107
The variable heavy chain comprising the amino acid sequence of SEQ ID NO: 14, which is 100% identical to the claimed SEQ ID NO: 30 as per claim 27, see col. 14, Table 3, reference SEQ ID NO: 14, sequence alignment below:
Query Match 100.0%; Score 617; Length 116;
Best Local Similarity 100.0%;
Matches 116; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
Qy 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSS 116
||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSS 116
Regarding claim 28, the ‘800 patent further teaches anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 47, which is identical to the claimed SEQ ID NO:
SEQ ID NO: 63, see col. 22, Table 7, reference SEQ ID NO: 47, and sequence alignment below:
Query Match 100.0%; Score 1121; Length 214;
Best Local Similarity 100.0%;
Matches 214; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
Qy 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIKRTVAAPSVFIFPP 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIKRTVAAPSVFIFPP 120
Qy 121 SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT 180
Qy 181 LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 214
||||||||||||||||||||||||||||||||||
Db 181 LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 214
and a heavy chain comprising the amino acid sequence of SEQ ID NO: 32, which is identical to the claimed SEQ ID NO: 48, see col. 19, Table 6, reference SEQ ID NO: 32, and sequence alignment below:
Query Match 100.0%; Score 2380; Length 445;
Best Local Similarity 100.0%;
Matches 445; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
Qy 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSSASTK 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSSASTK 120
Qy 121 GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS 180
Qy 181 LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVF 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVF 240
Qy 241 LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR 300
Qy 301 VVSVLTVLHQDWLNGKEYKCKVSNKARPAPIEKTISKAKGQPREPQVYTLPPSREEMTKN 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 VVSVLTVLHQDWLNGKEYKCKVSNKARPAPIEKTISKAKGQPREPQVYTLPPSREEMTKN 360
Qy 361 QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN 420
Qy 421 VFSCSVMHEALHNHYTQKSLSLSPG 445
|||||||||||||||||||||||||
Db 421 VFSCSVMHEALHNHYTQKSLSLSPG 445
Regarding claim 27, the ‘800 patent teaches anti-TfR1 antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region of SEQ ID NO 16, which comprises the claimed
HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19, see Table 15, sequence alignment below:
Query Match 87.1%; Score 166.4; Length 116;
Best Local Similarity 41.8%;
Matches 33; Conservative 0; Mismatches 0; Indels 46; Gaps 2;
Qy 1 YTFTNYWMH--------------EINPINGRSNYAEKFQG-------------------- 26
||||||||| |||||||||||||||||
Db 27 YTFTNYWMHWVRQAPGQGLEWMGEINPINGRSNYAEKFQGRVTLTVDTSSSTAYMELSSL 86
Qy 27 ------------GTRAMHY 33
|||||||
Db 87 RSEDTATYYCARGTRAMHY 105
and wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain (VL) region of SEQ ID NO: 22, which comprises the claimed LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, see Table 15, sequence alignment below:
Query Match 83.1%; Score 121.3; Length 107;
Best Local Similarity 36.5%;
Matches 27; Conservative 0; Mismatches 0; Indels 47; Gaps 2;
Qy 1 RTSENIYNNLA---------------AATNLAD--------------------------- 18
||||||||||| |||||||
Db 24 RTSENIYNNLAWYQQKQGKSPQLLVYAATNLADGVPSRFSGSGSGTQYSLKINSLQSEDF 83
Qy 19 -----QHFWGTPLT 27
|||||||||
Db 84 GNYYCQHFWGTPLT 97
The anti-transferrin receptor antibody has an improved serum half-life compared to other anti-transferrin receptor antibody, see col. 22, line 40-42. The anti-transferrin receptor antibody is used to conjugate any payload, e.g., PMO or siRNA, see col. 3, lines 23-34, Example 2, in particular.
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 combine the teachings of Levin and the ‘800 patent by substituting a known anti-TfR1 antibody or antigen binding fragment thereof in the conjugate comprising PMO of Levin for another, e.g., another the ‘800 patent’s anti-TfR1 antibody or antigen binding fragment thereof to arrive at the claimed invention with a reasonable expectation of success, preparing anti-TfR1 antibody-PMO for targeting PMO to the intracellular compartment of muscle cells.
One of ordinary skill in the art would have been motivated to do so because the ‘800 patent teaches that the anti-transferrin receptor antibody has an improved serum half-life compared to a reference anti-transferrin receptor antibody by at least 30 minutes to 7 days, see col. 22, line 40-42.
Further, “The substitution of familiar elements according to known methods is likely to be obvious when it does no more than yield predictable results.” 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.
Claims 16 and 23 are rejected under 35 U.S.C. 103 as being unpatentable over Levin et al (US20180369400, published December 27, 2018; PTO 892) in view of US patent No. 10,913,800 (issued February 9, 2021; PTO 892) as applied to claims 15 and 27-29 above and further in view of WO2019060775 publication (published March 28, 2019; PTO 892).
The combine teachings of Levin and the ‘800 patent have been discussed supra.
The references above do not teach the PMO comprises the sequence of SEQ ID NO:118 as per claim 16 and about 2.25 to 4.75 molar equivalents of the PMO conjugated with a linker are added in step c).
However, the WO2019060775 publication teaches a polynucleic acid conjugate comprising a target cell binding moiety binding to at least one polynucleic acid molecule that hybridizes to a target region of a pre-mRNA transcript of DMD gene, wherein the at least one polynucleic acid molecule induces splicing out of an exon from a pre-mRNA transcript to generate a mRNA transcript that encodes a functional dystrophin protein, see entire document, abstract, p. 1, Summary of disclosure. The reference oligonucleotide comprises the sequence of SEQ ID NO: 1063, which is 100% identical to the claimed SEQ ID NO: 118, see p. 134, reference SEQ ID NO 1063, sequence alignment below:
Query Match 100.0%; Score 26; Length 30;
Best Local Similarity 100.0%;
Matches 26; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CGCCGCCATTTCTCAACAGATCTGTC 26
||||||||||||||||||||||||||
Db 5 CGCCGCCATTTCTCAACAGATCTGTC 30
The term “comprises” is open ended. It expands the claimed oligonucleotide to include additional residues at the 5’ end.
The reference oligonucleotide is a phosphorodiamidate morpholino oligomer (PMO) sequence, see Fig. 1, in particular. Example of target cell binding moiety includes anti-transferrin receptor antibody (aka anti-CD71, see para. [0407] to [0408]. The WO2019060775 publication teaches Anti-CD71 antibody morpholino antisense oligonucleotide conjugate (anti-CD71 mAb-PMO), see p. 89 for linker, p. 404, para. [0409] to [0410].
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 conjugate Levin and the ‘800 patent’s anti-transferrin receptor antibody to WO2019060775 publication’s phosphorodiamidate morpholino oligomer (PMO) sequence that is identical to the claimed SEQ ID NO: 118 in order to form Anti-CD71 mAb-PMO conjugation for targeting PMO to muscle cells expressing transferrin receptor for internalization.
One of ordinary skill in the art would have had a reasonable expectation of success at the time the invention was made to modify the method of Levin and the ‘800 patent in view of WO2019060775 publication because the WO2019060775 publication teaches that the PMO hybridizes to a target region of a pre-mRNA transcript of DMD gene and induces splicing out of an exon 44 from a pre-mRNA transcript to generate a mRNA transcript that encodes a functional dystrophin protein and conjugating PMO to anti-transferrin receptor antibody is expected to bind to human transferrin receptor for receptor mediated uptake and internalization of PMO into muscle cells (see para. [0438], [0466]).
One of ordinary skill in the art would have been motivated to do so because the WO2019060775 publication teaches that the anti-CD71 (aka transferrin receptor) mAb-PMO is useful for treatment of a muscular dystrophy, and transferrin receptor is abundant on muscle cells, making it a target for drug delivery, see Summary of invention.
One of ordinary skill in the art would have been motivated to do so because the ‘800 patent teaches that the anti-transferrin receptor antibody described herein has an improved serum half-life compared to a reference anti-transferrin receptor antibody by at least 30 minutes to 7 days, see col. 22, line 40-42.
Further, using a known method of conjugating anti-transferrin antibody to PMO of Levin or the WO2019060775 publication to a known anti-transferrin receptor antibody of the ‘800 patent to a known PMO comprising SEQ ID NO: 118 of WO2019060775 publication would improve similar products in the same way.
In the alternative, simple substitution of a known element, anti-TfR antibody in the method of WO2019060775 publication for the known anti-TfR antibody of the ‘800 patent would obtain predictable results. KSR Int’l Co. v. Teleflex Inc., 550 U.S. 398, 416 (2007).
Claim 23 is included because it would have been obvious to one of ordinary skill in the art at the time of the invention to engage in routine experimentation to determine optimal or workable ranges, e.g., about 2.25 to 4.75 molar equivalents of PMO that produce expected results. Where the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation. In re Alter, 220 F. 2d 454, 105 USPQ 233 (CCPA 1955). NOTE: MPEP 2144.05.
A person of ordinary skill in the art is always motivated to pursue the known options within her or his technical grasp. If this leads to the anticipated success, it is likely the product not of innovation but of ordinary skill and common sense.
“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.
Claims 1, 3-4 and 6-12 are rejected under 35 U.S.C. 103 as being unpatentable over Levin et al (US20180369400, published December 27, 2018; PTO 892) in view of Alley et al (US20090010945, published January 8, 2009; PTO 892), Hanson et al (US20120065169, published March 15, 2012; PTO 892) and Scheer (US20110287009, published November 24, 2011; PTO 892).
Claim 1 recites a method of preparing an anti-transferrin receptor (TfR1) antibody- PMO conjugate comprising the steps of:
a) contacting an anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof with a reducing agent thereby producing a reduced anti-TfR1 antibody or a reduced anti-TfR1 antigen binding fragment thereof;
b) purifying the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof;
c) contacting a PMO with a linker thereby generating a PMO-linker;
d) contacting the PMO-linker with the reduced anti-TfR1 antibody or the reduced anti-TfR1 antigen binding fragment thereof, thereby generating an anti-TfR1 antibody-PMO conjugate or an anti-TfR1 antigen binding fragment-PMO conjugate;
e) capping the anti-TfR1 antibody or the anti-TfR1 antigen binding fragment thereof using an alkylating agent; and
f) isolating the anti-TfR1 antibody-PMO conjugate or anti-TfR1 antigen binding fragment-PMO conjugate, thereby preparing the anti-TfR1 antibody-PMO conjugate or the anti-TfR1 antigen binding fragment-PMO conjugate.
Claim 3 recites the method of claim 1, wherein the reducing agent comprises tris(2- carboxyethyl)phosphine (TCEP).
Claim 4 recites the method of claim 3, wherein about 2 to 4 molar equivalents of TCEP are used in step a).
Claim 6 recites the method of claim 1, wherein the linker is attached at the 3' terminus of the PMO.
Claim 7 recites the method of claim 1, wherein the alkylating agent comprises N-ethylmaleimide (NEM) and the linker comprises a maleimide group.
Claim 8 recites the method of claim 1, wherein the linker comprises SMCC (elected specie).
Claim 9 encompasses the method of claim 1, wherein about 2.25 to 4.75 molar equivalents of PMO- linker is used in step d) and about 7.5 to 10 molar equivalents of the alkylating agent comprising NEM is used are added in step e).
Claim 10 encompasses the method of claim 1, wherein the reduced anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof is purified by filtration or chromatography (elected species).
Claim 12 encompasses the method of claim 1, wherein the isolated anti-TfR1 antibody-PMO conjugate or the anti-TfR1 antigen binding fragment-PMO conjugate has an average DAR ratio of about 3.5-4.5.
Regarding claims 1, 3, 6, 7, 8, 10, Levin teaches a method of preparing an anti-transferrin receptor (TfR1) antibody-PMO conjugate, the reference method comprises: a) contacting anti-CD71 (aka transferrin receptor (TfR1) antibody or antigen-binding fragment thereof, e.g., F(ab’)2 ([0487]) with a reducing agent by adding 4 equivalents tris(2-carboxyethyl)phosphine (TCEP) to produce a reduced anti-CD71 antibody or antigen-binding fragment thereof as per step a in claim 1, b) contacting PMO with a linker, e.g., 4(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) to generate PMO-SMCC as per step c in claim 1, c) incubate the reduced antibody and PMO-SMCC to generate antibody-PMO conjugates as per step d in claim 1, d) capping the anti-TfR1 antibody or antigen-binding fragment thereof using 8 equivalents of an alkylating agent, e.g., N-Ethylmaleimide (NEM) to quench unreacted cysteines (aka capping the cysteines on the anti-CD71 antibody or antigen-binding fragment thereof by modify sulfhydryl groups as per step e in claim 1, see para. [0480] and then isolating the anti-CD71 mAb-PMO by hydrophobic interaction chromatography (HIC) as per step f in claim 1, see para. [0477] to [0480]. Levin teaches that that purification methods include size exclusion chromatography, see Table 1, hydrophobic interaction chromatograph (HIC, Tables 6-9), strong anion exchange chromatography (SAX, Table 10-11). Levin teaches that the reaction mixture can be purified using the first Protein Liquid Chromatography (FPLC), see para. [0488].
Regarding claim 4, Levin teaches 4 molar equivalents of reducing agent, tris(2-carboxyethyl)phosphine (TCEP) was used in step a, see para. [0480].
Regarding claim 6, Levin teaches that the SMCC was coupled to the primary amine on the 3′ end of the phosphorodiamidate morpholino oligomer (PMO), see para. [0480].
Regarding claim 9, Levin teaches that the 8 equivalent of N-Ethylmaleimide (NEM), which is a potent alkylating agent primarily to modify sulfhydryl group. The reference 8 molar equivalents is within the claimed range of about 7.5 to 10 MEM. It would have been obvious to one of ordinary skill in the art at the time of the invention to engage in routine experimentation to determine optimal or workable ranges, e.g., about 2.25 to 4.75 molar equivalents of PMO that produce expected results. Where the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation. In re Alter, 220 F. 2d 454, 105 USPQ 233 (CCPA 1955). NOTE: MPEP 2144.05.
Regarding claim 12, Levin teaches that the drug-to-antibody ratio is about 3, 4, 5 or greater, see para. [0342] to [0344].
Levin does not teach the method purifying the reduced anti-TfR1 antibody or antigen fragment thereof before contacting PMO-linker in step b in claim 1, and the reduced antibody or antigen-binding fragment thereof is purified by strong cation exchange chromatography (SCX) as per claim 11.
However, Alley teaches a method of producing antibody-drug conjugate (ADCs, see entire document, para. [0006], [0007]). The method generally includes fully reducing the antibody interchain disulfide bonds with a reducing agent, such as but not limited to dithiothreitol (DTT), Tris(2-carboxyethyl)phosphine (TCEP), or 2-Mercaptoethanol, breaks all four bonds and generates eight cysteines (i.e., containing the free thiol group). Conjugation of all eight cysteines with the drug-linker generates a fully-loaded conjugate with approximately eight drugs per antibody, as shown in see FIG. 7, see para. [0085]. The reduced antibody can be purified prior to conjugation, using for example, column chromatography, dialysis, or diafiltration. The column used in column chromatography can be, for example, a desalting column, such as a PD-10 column. Alternatively, the reduced antibody is not purified after partial reduction and prior to conjugation, see para. [0020]. To partially reduce the antibody, 2.20 molar equivalents of TCEP is added to the antibody solution and allowed to reduce for 105 min at 37.degree. C, see para. [0033]. Alley teaches a slight excess of the drug-linker vcMMAE (typically 2-15% excess in the form of a DMSO solution) is then added into the antibody solution to start the conjugation reaction, see para. [0033].
Alley does not teach strong Cation Exchange Chromatography to purified reduced antibody.
However, Hanson teaches a method of purifying phosphorodiamidate morpholino oligomers (morpholino oligomers, instant PMO) using Cation Exchange Chromatography, see para. [0499].
Scheer teaches the use of strong Cation Exchange Chromatography to purified reduced antibody (aka half antibody, see para. [0392]. Scheer teaches that protein purification such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE.TM. chromatography on an anion or cation exchange resin (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available depending on the antibody to be recoveredmm, see para. [0294].
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 purify Levin’s reduced antibody or morpholino oligomers (PMO) using strong Cation Exchange Chromatography as taught by Scheer or Hanson prior to contacting the reduced antibody with the drug-linker as taught by Alley in order to remove any excess reducing agent and aggregates based on charge prior to contacting with the PMO-SMCC, thereby increasing the yield of the final anti-PMO conjugates.
One of ordinary skill in the art would have been motivated to purifying the reduced antibody or antigen-binding fragment thereof because Alley teaches that it is typically purifying reduced antibody prior to conjugating the drug linker, see para. [0020].
One of ordinary skill in the art would have been motivated to use cation exchange chromatography because Hanson teaches that PMO can be purified using Cation Exchange Chromatography (see para. [0499]) and Scheer teaches that antibody purification using strong Cation Exchange Chromatography is known in the art.
A person of ordinary skill in the art is always motivated to pursue the known options within her or his technical grasp. If this leads to the anticipated success, it is likely the product not of innovation but of ordinary skill and common sense.
“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.
Claims 2 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Levin et al (US20180369400, published December 27, 2018; PTO 892) in view of Hanson et al (US20120065169, published March 15, 2012; PTO 892) and Scheer (US20110287009, published November 24, 2011; PTO 892) as applied to claims 1, 3-4 and 6-12 and further in view of WO2019060775 publication (published March 28, 2019; PTO 892).
The teachings of Levin, Hanson and Scheer have been discussed supra.
The references do not teach the method of preparing anti-TfR1 antibody-PMO conjugate wherein the PMO comprises the sequence of SEQ ID NO: 118 as per claim 2, wherein about 3 molar equivalent of linker are used in step c as per claim 5.
However, the WO2019060775 publication teaches a polynucleic acid conjugate comprising a target cell binding moiety binding to at least one polynucleic acid molecule that hybridizes to a target region of a pre-mRNA transcript of DMD gene, wherein the at least one polynucleic acid molecule induces splicing out of an exon from a pre-mRNA transcript to generate a mRNA transcript that encodes a functional dystrophin protein, see entire document, abstract, p. 1, Summary of disclosure. The reference oligonucleotide comprises the sequence of SEQ ID NO: 1063, which is 100% identical to the claimed SEQ ID NO: 118, see p. 134, reference SEQ ID NO 1063, sequence alignment below:
Query Match 100.0%; Score 26; Length 30;
Best Local Similarity 100.0%;
Matches 26; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CGCCGCCATTTCTCAACAGATCTGTC 26
||||||||||||||||||||||||||
Db 5 CGCCGCCATTTCTCAACAGATCTGTC 30
The term “comprises” is open ended. It expands the claimed oligonucleotide to include additional residues at the 5’ end.
The reference oligonucleotide is a phosphorodiamidate morpholino oligomer (PMO) sequence, see Fig. 1, in particular. Example of target cell binding moiety includes anti-transferrin receptor antibody (aka anti-CD71, see para. [0407] to [0408]. The WO2019060775 publication teaches Anti-CD71 antibody morpholino antisense oligonucleotide conjugate (anti-CD71 mAb-PMO), see p. 89 for linker, p. 404, para. [0409] to [0410].
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 conjugate Levin, Hanson and Scheer’s anti-transferrin receptor antibody to WO2019060775 publication’s phosphorodiamidate morpholino oligomer (PMO) sequence that is identical to the claimed SEQ ID NO: 118 in order to form Anti-CD71 mAb-PMO conjugation for targeting PMO to muscle cells expressing transferrin receptor for internalization.
One of ordinary skill in the art would have had a reasonable expectation of success at the time the invention was made to modify the method of Levin, Hanson and Scheer in view of WO2019060775 publication because the WO2019060775 publication teaches that the PMO hybridizes to a target region of a pre-mRNA transcript of DMD gene and induces splicing out of an exon 44 from a pre-mRNA transcript to generate a mRNA transcript that encodes a functional dystrophin protein and then conjugating PMO to anti-transferrin receptor antibody is expected to bind to human transferrin receptor for receptor mediated uptake and internalization of PMO into muscle cells (see para. [0438], [0466]).
One of ordinary skill in the art would have been motivated to do so because the WO2019060775 publication teaches that the anti-CD71 (aka transferrin receptor) mAb-PMO is useful for treatment of a muscular dystrophy, and transferrin receptor is abundant on muscle cells, making it a target for drug delivery, see Summary of invention.
Further, using a known method of conjugating anti-transferrin antibody to PMO of Levin or the WO2019060775 publication to a known PMO comprising SEQ ID NO: 118 of WO2019060775 publication would improve similar products in the same way. KSR Int’l Co. v. Teleflex Inc., 550 U.S. 398, 416 (2007).
Claim 5 is included because it would have been obvious to one of ordinary skill in the art at the time of the invention to engage in routine experimentation to determine optimal or workable ranges, e.g., about 2.25 to 4.75 molar equivalents of PMO that produce expected results. Where the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation. In re Alter, 220 F. 2d 454, 105 USPQ 233 (CCPA 1955). NOTE: MPEP 2144.05.
A person of ordinary skill in the art is always motivated to pursue the known options within her or his technical grasp. If this leads to the anticipated success, it is likely the product not of innovation but of ordinary skill and common sense.
“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.
Claims 13-14 and 30 are rejected under 35 U.S.C. 103 as being unpatentable over Levin et al (US20180369400, published December 27, 2018; PTO 892) in view of Hanson et al (US20120065169, published March 15, 2012; PTO 892) and Scheer (US20110287009, published November 24, 2011; PTO 892) as applied to claims 1, 3-4 and 6-12 and further in view of US patent No. 10,913,800 (issued February 9, 2021; PTO 892).
The teachings of Levin, Hanson and Scheer have been discussed supra.
The references do not teach the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable light chain comprising the amino acid sequence of SEQ ID NO: 34 and a variable heavy chain comprising the amino acid sequence of SEQ ID NO: 30 as per claim 13 or wherein the anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 63 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 as per claim 14 or wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19; and wherein the anti-TfR1 antibody or anti-TfR1 antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24 as per claim 30.
However, the ‘800 patent teaches anti-TfR1 antibody or antigen comprising and a light variable heavy chain comprising the amino acid sequence of SEQ ID NO: 18, which is identical to the claimed SEQ ID NO: 34, see col. 14, Table 14, reference SEQ ID NO: 18, 3 CDRs are underline, sequence alignment below:
Query Match 100.0%; Score 568; Length 107;
Best Local Similarity 100.0%;
Matches 107; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
Qy 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIK 107
|||||||||||||||||||||||||||||||||||||||||||||||
Db 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIK 107
The variable heavy chain comprising the amino acid sequence of SEQ ID NO: 14, which is identical to the claimed SEQ ID NO: 30 as per claim 27, see col. 14, Table 3, reference SEQ ID NO: 14, sequence alignment below:
Query Match 100.0%; Score 617; Length 116;
Best Local Similarity 100.0%;
Matches 116; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
Qy 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSS 116
||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSS 116
Regarding claim 14, the ‘800 patent further teaches anti-TfR1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 47, which is identical to the claimed SEQ ID NO:
SEQ ID NO: 63, see col. 22, Table 7, reference SEQ ID NO: 47, and sequence alignment below:
Query Match 100.0%; Score 1121; Length 214;
Best Local Similarity 100.0%;
Matches 214; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 DIQMTQSPSSLSASVGDRVTITCRTSENIYNNLAWYQQKPGKSPKLLIYAATNLADGVPS 60
Qy 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIKRTVAAPSVFIFPP 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 RFSGSGSGTDYTLTISSLQPEDFATYYCQHFWGTPLTFGGGTKVEIKRTVAAPSVFIFPP 120
Qy 121 SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT 180
Qy 181 LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 214
||||||||||||||||||||||||||||||||||
Db 181 LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 214
and a heavy chain comprising the amino acid sequence of SEQ ID NO: 32, which is identical to the claimed SEQ ID NO: 48, see col. 19, Table 6, reference SEQ ID NO: 32, and sequence alignment below:
Query Match 100.0%; Score 2380; Length 445;
Best Local Similarity 100.0%;
Matches 445; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMHWVRQAPGQGLEWIGEINPINGRSNY 60
Qy 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSSASTK 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 AEKFQGRVTLTVDTSSSTAYMELSRLRSDDTAVYYCARGTRAMHYWGQGTLVTVSSASTK 120
Qy 121 GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS 180
Qy 181 LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVF 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVF 240
Qy 241 LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR 300
Qy 301 VVSVLTVLHQDWLNGKEYKCKVSNKARPAPIEKTISKAKGQPREPQVYTLPPSREEMTKN 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 VVSVLTVLHQDWLNGKEYKCKVSNKARPAPIEKTISKAKGQPREPQVYTLPPSREEMTKN 360
Qy 361 QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN 420
Qy 421 VFSCSVMHEALHNHYTQKSLSLSPG 445
|||||||||||||||||||||||||
Db 421 VFSCSVMHEALHNHYTQKSLSLSPG 445
Regarding claim 30, the ‘800 patent teaches anti-TfR1 antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region of SEQ ID NO 16, which comprises the claimed
HCDR1 comprising the amino acid sequence of SEQ ID NO: 17; an HCDR2 comprising the amino acid sequence of SEQ ID NO: 20; and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 19, see Table 15, sequence alignment below:
Query Match 87.1%; Score 166.4; Length 116;
Best Local Similarity 41.8%;
Matches 33; Conservative 0; Mismatches 0; Indels 46; Gaps 2;
Qy 1 YTFTNYWMH--------------EINPINGRSNYAEKFQG-------------------- 26
||||||||| |||||||||||||||||
Db 27 YTFTNYWMHWVRQAPGQGLEWMGEINPINGRSNYAEKFQGRVTLTVDTSSSTAYMELSSL 86
Qy 27 ------------GTRAMHY 33
|||||||
Db 87 RSEDTATYYCARGTRAMHY 105
and wherein the anti-TfR1 antibody or antigen binding fragment thereof comprises a variable light chain (VL) region of SEQ ID NO: 22, which comprises the claimed LCDR1 comprising the amino acid sequence of SEQ ID NO: 22; an LCDR2 comprising the amino acid sequence of SEQ ID NO: 23; and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 24, see Table 15, sequence alignment below:
Query Match 83.1%; Score 121.3; Length 107;
Best Local Similarity 36.5%;
Matches 27; Conservative 0; Mismatches 0; Indels 47; Gaps 2;
Qy 1 RTSENIYNNLA---------------AATNLAD--------------------------- 18
||||||||||| |||||||
Db 24 RTSENIYNNLAWYQQKQGKSPQLLVYAATNLADGVPSRFSGSGSGTQYSLKINSLQSEDF 83
Qy 19 -----QHFWGTPLT 27
|||||||||
Db 84 GNYYCQHFWGTPLT 97
The anti-transferrin receptor antibody has an improved serum half-life compared to other anti-transferrin receptor antibody, see col. 22, line 40-42. The anti-transferrin receptor antibody is used to conjugate any payload, e.g., PMO or siRNA, see col. 3, lines 23-34, Example 2, 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 combine the teachings of Levin, Hanson and Scheer in view the ‘800 patent by substituting a known anti-TfR1 antibody or antigen binding fragment thereof in the conjugate comprising PMO for another, e.g., another anti-TfR1 antibody or antigen binding fragment thereof to arrive at the claimed invention with a reasonable expectation of success, targeting PMO to the intracellular compartment and improved serum half-life of the conjugate.
One of ordinary skill in the art would have been motivated to do so because the ‘800 patent teaches that the anti-transferrin receptor antibody described herein has an improved serum half-life compared to a reference anti-transferrin receptor antibody by at least 30 minutes to 7 days, see col. 22, line 40-42.
Further, “The substitution of familiar elements according to known methods is likely to be obvious when it does no more than yield predictable results.” 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.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the claims at issue are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); and In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on a nonstatutory double patenting ground provided the reference application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The USPTO internet Web site contains terminal disclaimer forms which may be used. Please visit http://www.uspto.gov/forms/. The filing date of the application will determine what form should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to http://www.uspto.gov/patents/process/file/efs/guidance/eTD-info-I.jsp.
Claims 1-30 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-18 of copending Application No. 19/234,136 (reference). Although the claims at issue are not identical, they are not patentably distinct from each other because while the ‘136 reference application which served as the parent for the present case, the examined application was filed as a CON, not a DIV, and therefore no shield against double patenting that might be provided by 35 U.S.C 121 would be applicable here.
Copending claims recite:
1. (Original) A conjugate comprising (i) an anti-transferrin receptor antibody or antigen binding fragment thereof, (ii) a phosphorodiamidate morpholino oligonucleotide (PMO), and (iii) a linker, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) region, which comprises an HCDR1 comprising the sequence of SEQ ID NO: 17; an HCDR2 comprising the sequence of SEQ ID NO: 20; and an HCDR3 comprising the sequence of SEQ ID NO: 19;wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a variable light chain (VL) region, which comprises a LCDR1 comprising the sequence of SEQ ID NO: 22; a LCDR2 comprising the sequence of SEQ ID NO: 23; and a LCDR3 comprising the sequence of SEQ ID NO: 24;wherein the PMO comprises the sequence of SEQ ID NO: 118; and wherein the linker comprises a maleimide group that conjugates the anti-transferrin receptor antibody or antigen binding fragment thereof to a terminus of the PMO, which corresponds to instant claims 29-30.
2. (Original) The conjugate of claim 1, wherein the maleimide group is selected from the group consisting of succinimidyl 4-(N-maleimidomethyl)cyclohexane-l-carboxylate (SMCC) and sulfosuccinimidyl-4-(N-maleimidomethyl)cycloexane-l-carboxylate (sulfo-sMCC), which corresponds to instant claims 7, 8, 21.
3. (Original) The conjugate of claim 1, wherein the linker conjugates the anti-transferrin receptor antibody or antigen binding fragment thereof to the 3' end terminus of the PMO, which corresponds to instant claims 1, 15.
4. (Original) The conjugate of claim 1, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof is a full-length antibody.
5. (Original) The conjugate of claim 4, wherein the full-length antibody is a humanized antibody or a human antibody.
6. (Original) The conjugate of claim 4, wherein the full-length anti-transferrin receptor antibody further comprises a mutation in the heavy chain constant region selected from the group consisting of L233A, L234A, and L327R, which corresponds to instant claims 14 and 28.
7. (Original) The conjugate of claim 6, wherein the full-length anti-transferrin receptor antibody further comprises the L233A, L234A and L327R mutations in the heavy chain constant region.
8. (Original) The conjugate of claim 1, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof is selected from the group consisting of monovalent Fab', divalent Fab2, and single chain variable fragment (scFv).
9. (Original) A conjugate comprising (i) an anti-transferrin receptor antibody or antigen binding fragment thereof, (ii) a phosphorodiamidate morpholino oligonucleotide (PMO), and (iii) a linker, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a variable heavy chain (VH) sequence of SEQ ID NO: 30 and a variable light chain (VL) sequence of SEQ ID NO: 34;wherein the PMO comprises the sequence of SEQ ID NO: 118; and wherein the linker comprises a maleimide group that conjugates the anti-transferrin receptor antibody or antigen binding fragment thereof to a terminus of the PMO.
10. (Original) The conjugate of claim 9, wherein the maleimide group is selected from the group consisting of succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) and sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-sMCC).
11. (Original) The conjugate of claim 9, wherein the linker conjugates the anti-transferrin antibody or antigen binding fragment thereof to the 3' end terminus of the PMO.
12. (Original) The conjugate of claim 9, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof is a full-length anti-transferrin receptor antibody.
13. (Original) The conjugate of claim 12, wherein the full-length anti-transferrin receptor antibody is a humanized anti-transferrin receptor antibody or a human anti-transferrin receptor antibody.
14. (Original) The conjugate of claim 12, wherein the full-length anti-transferrin receptor antibody further comprises a mutation in the heavy chain constant region selected from the group consisting of L233A, L234A, and L327R.
15. (Original) The conjugate of claim 14, wherein the full-length anti-transferrin receptor antibody further comprises the L233A, L234A and L327R mutations in the heavy chain constant region.
16. (Original) The conjugate of claim 9, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof is selected from the group consisting of monovalent Fab', divalent Fab2, and single chain variable fragment (scFv).
17. (Original) A conjugate comprising (i) an anti-transferrin receptor antibody, (ii) a phosphorodiamidate morpholino oligonucleotide (PMO), and (iii) a linker, wherein the anti-transferrin receptor antibody comprises (a) two heavy chains, wherein each of the two heavy chains comprises SEQ ID NO: 48, and (b) two light chains, wherein each of the two light chains comprises SEQ ID NO: 63;wherein the PMO comprises the sequence of SEQ ID NO: 118; and wherein the linker comprises a maleimide group that conjugates the anti-transferrin receptor antibody to the 3' terminus of the PMO.
18. (Original) The conjugate of claim 17, wherein the maleimide group is selected from the group consisting of succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) and sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-sMCC).
19/329,403
19/234,136
SEQ ID NO: 118
cgccgccatttctcaacagatctgtc
cgccgccatttctcaacagatctgtc
SEQ ID NO: 17
YTFTNYWMH
YTFTNYWMH
SEQ ID NO: 20
EINPINGRSNYAEKFQG
EINPINGRSNYAEKFQG
SEQ ID NO: 19
GTRAMHY
GTRAMHY
SEQ ID NO: 22
RTSENIYNNLA
RTSENIYNNLA
SEQ ID NO: 23
AATNLAD
AATNLAD
SEQ ID NO: 24
QHFWGTPLT
QHFWGTPLT
SEQ ID NO: 34
SEQ ID NO: 34
SEQ ID NO: 30
SEQ ID NO: 30
SEQ ID NO: 63
SEQ ID NO: 63
SEQ ID NO: 48
SEQ ID NO: 48
A person of skill in the art, reading the claims of the ‘136 application, would look to the application and follow the ‘136 application’s express instruction on how to produce the product using the method in the ‘136 application, e.g., Example 7 at p. 91-93, thereby arriving at the claimed method of the examined claims.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Claims 1- 30 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-21 of U.S. Patent No. 12,359,202. Although the claims at issue are not identical, they are not patentably distinct from each other because while the ‘342 patent issued from the application which served as the grandparent for the present case, the examined application was filed as a CON, not a DIV, and therefore no shield against double patenting that might be provided by 35 U.S.C 121 would be applicable here.
Issued claim:
1. An antisense oligonucleotide conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to an antisense oligonucleotide, wherein the antisense oligonucleotide consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 116-119.
2. The antisense oligonucleotide conjugate of claim 1, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a humanized antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, monoclonal antibody or antigen binding fragment thereof, monovalent Fab′, divalent Fab.sub.2, single chain variable fragment (scFv), diabody, minibody, nanobody, single domain antibody (sdAb), or camelid antibody or antigen binding fragment thereof.
3. The antisense oligonucleotide conjugate of claim 1, wherein the antisense oligonucleotide is conjugated to the anti-transferrin receptor antibody or antigen binding fragment thereof via a linker.
4. The antisense oligonucleotide conjugate of claim 3, wherein the linker is a cleavable linker or a non-cleavable linker, wherein the linker is a heterobifunctional linker or a homobifunctional linker, and wherein the linker comprises a maleimide group, a dipeptide moiety, a benzoic acid group or derivatives thereof, a C1-C6 alkyl group, or a combination thereof.
5. The antisense oligonucleotide conjugate of claim 1, wherein the antisense oligonucleotide conjugate has a ratio of the antisense oligonucleotide and the anti-transferrin receptor antibody or antigen binding fragment thereof as 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or higher.
6. The antisense oligonucleotide conjugate of claim 5, wherein the antisense oligonucleotide conjugate has the ratio as 4 or 8 on average.
7. A method of treating muscular dystrophy in a subject in need thereof comprising administering to the subject an antisense oligonucleotide conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to an antisense oligonucleotide, wherein the antisense oligonucleotide consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 116-119, wherein the antisense oligonucleotide induces exon 44 skipping in a pre-mRNA transcript of a DMD gene to generate an mRNA transcript encoding a truncated dystrophin protein.
8. The method of claim 7, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a humanized antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, monoclonal antibody or antigen binding fragment thereof, monovalent Fab′, divalent Fab.sub.2, single chain variable fragment (scFv), diabody, minibody, nanobody, single domain antibody (sdAb), or camelid antibody or antigen binding fragment thereof.
9. The method of claim 7, wherein the antisense oligonucleotide is conjugated to the anti-transferrin receptor antibody or antigen binding fragment thereof via a linker.
10. The method of claim 9, wherein the linker is a cleavable linker or a non-cleavable linker, wherein the linker is a heterobifunctional linker or a homobifunctional linker, and wherein the linker comprises a maleimide group, a dipeptide moiety, a benzoic acid group or derivatives thereof, a C1-C6 alkyl group, or a combination thereof.
11. The method of claim 7, wherein the antisense oligonucleotide conjugate has a ratio of the antisense oligonucleotide and the anti-transferrin receptor antibody or antigen binding fragment thereof as 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or higher.
12. The method of claim 11, wherein the antisense oligonucleotide conjugate has the ratio as 4 or 8 on average.
13. The method of claim 7, wherein the antisense oligonucleotide conjugate is administered parenterally.
14. The method of claim 7, wherein the truncated dystrophin protein modulates muscular dystrophy.
15. The method of claim 7, wherein the muscular dystrophy is Duchenne muscular dystrophy or Becker muscular dystrophy.
16. A method of inducing exon 44 skipping in a targeted pre-mRNA transcript of a DMD gene, comprising: a) contacting a muscle cell with an antisense oligonucleotide conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to an antisense oligonucleotide, wherein the antisense oligonucleotide consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 116-119, wherein the antisense oligonucleotide induces exon 44 skipping in the targeted pre-mRNA transcript, and wherein the antisense oligonucleotide conjugate is preferentially delivered into the muscle cell; b) hybridizing the antisense oligonucleotide to the targeted pre-mRNA transcript to induce exon 44 skipping in the targeted pre-mRNA transcript; and c) translating an mRNA transcript produced from the targeted pre-mRNA transcript processed in step b) in the muscle cell to generate a truncated dystrophin protein.
17. The method of claim 16, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a humanized antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, monoclonal antibody or antigen binding fragment thereof, monovalent Fab′, divalent Fab.sub.2, single chain variable fragment (scFv), diabody, minibody, nanobody, single domain antibody (sdAb), or camelid antibody or antigen binding fragment thereof.
18. The method of claim 16, wherein the antisense oligonucleotide is conjugated to the anti-transferrin receptor antibody or antigen binding fragment thereof via a linker.
19. The method of claim 18, wherein the linker is a cleavable linker or a non-cleavable linker, wherein the linker is a heterobifunctional linker or a homobifunctional linker, and wherein the linker comprises a maleimide group, a dipeptide moiety, a benzoic acid group or derivatives thereof, a C1-C6 alkyl group, or a combination thereof.
20. The method of claim 16, wherein the antisense oligonucleotide conjugate has a ratio of the antisense oligonucleotide and the anti-transferrin receptor antibody or antigen binding fragment thereof as 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or higher.
21. The method of claim 20, wherein the antisense oligonucleotide conjugate has the ratio as 4 or 8 on average.
19/329,403
12,359,202
SEQ ID NO: 118
cgccgccatttctcaacagatctgtc
cgccgccatttctcaacagatctgtc
SEQ ID NO: 17
YTFTNYWMH
YTFTNYWMH
SEQ ID NO: 20
EINPINGRSNYAEKFQG
EINPINGRSNYAEKFQG
SEQ ID NO: 19
GTRAMHY
GTRAMHY
SEQ ID NO: 22
RTSENIYNNLA
RTSENIYNNLA
SEQ ID NO: 23
AATNLAD
AATNLAD
SEQ ID NO: 24
QHFWGTPLT
QHFWGTPLT
SEQ ID NO: 34
SEQ ID NO: 34
SEQ ID NO: 30
SEQ ID NO: 30
SEQ ID NO: 63
SEQ ID NO: 63
SEQ ID NO: 48
SEQ ID NO: 48
A person of skill in the art, reading the claims of the ‘202 patent, would look to the patent and follow the ‘202 patent’s express instruction on how to make the product within the patent, e.g., Example 7, thereby arriving at the claimed method of the examined claims.
Claims 1-30 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-21 of U.S. Patent No. 12,071,621. Although the claims at issue are not identical, they are not patentably distinct from each other because while the ‘621 patent issued from the application which served as the great grandparent for the present case, the examined application was filed as a CON, not a DIV, and therefore no shield against double patenting that might be provided by 35 U.S.C 121 would be applicable here.
Issued claims:
1. A phosphorodiamidate morpholino oligonucleotide (PMO) conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to a PMO molecule, wherein the PMO molecule consists of a sequence selected from the group consisting of SEQ ID NOs: 116-119.
2. The PMO conjugate of claim 1, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a humanized antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, monoclonal antibody or antigen binding fragment thereof, monovalent Fab′, divalent Fab.sub.2, single chain variable fragment (scFv), diabody, minibody, nanobody, single domain antibody (sdAb), or camelid antibody or antigen binding fragment thereof. The claimed PMO of SEQ ID NO: 118 is the same as instant SEQ ID NO: 118 in instant claims 2 and 16.
3. The PMO conjugate of claim 1, wherein the PMO molecule is conjugated to the anti-transferrin receptor antibody or antigen binding fragment thereof via a linker.
4. The PMO conjugate of claim 3, wherein the linker is a cleavable linker or a non-cleavable linker, wherein the linker is a heterobifunctional linker or a homobifunctional linker, and wherein the linker comprises a maleimide group, a dipeptide moiety, a benzoic acid group or derivatives thereof, a C.sub.1-C.sub.6 alkyl group, or a combination thereof.
5. The PMO conjugate of claim 1, wherein the PMO conjugate has a PMO molecule to antibody ratio (DAR) of 1:1, 2:1, 3:1, 4:1% 5:1, 6:1, 7:1, 8:1 or higher.
6. The PMO conjugate of claim 1, wherein the PMO conjugate has an average DAR of 1, 2, 3, 4, 5, 6, 7, 8 or higher.
7. The PMO conjugate of claim 1, wherein the PMO conjugate has an average DAR in the range of 3.5-4.5 or 7.5-8.5.
8. The PMO conjugate of claim 1, wherein the PMO conjugate has an average DAR of 4 or 8.
9. The PMO conjugate of claim 1, wherein the PMO conjugate has a DAR of 4 or 8.
10. A method of treating muscular dystrophy in a subject in need thereof comprising administering to said subject a phosphorodiamidate morpholino oligonucleotide (PMO) conjugate comprising an anti-transferrin receptor antibody or antigen binding fragment thereof conjugated to a PMO molecule comprising a sequence selected from the group consisting of SEQ ID NOs: 116-119, wherein the PMO molecule induces exon 44 skipping in a pre-mRNA transcript of a DMD gene to generate an mRNA transcript encoding a truncated dystrophin protein.
11. The method of claim 10, wherein the PMO molecule is delivered into a muscle cell.
12. The method of claim 10, wherein the anti-transferrin receptor antibody or antigen binding fragment thereof comprises a humanized antibody or antigen binding fragment thereof, chimeric antibody or antigen binding fragment thereof, monoclonal antibody or antigen binding fragment thereof, monovalent Fab′, divalent Fab.sub.2, single chain variable fragment (scFv), diabody, minibody, nanobody, single domain antibody (sdAb), or camelid antibody or antigen binding fragment thereof.
13. The method of claim 10, wherein the PMO molecule is conjugated to the anti-transferrin receptor antibody or antigen binding fragment thereof via a linker.
14. The method of claim 13, wherein the linker is a cleavable linker or a non-cleavable linker, wherein the linker is a heterobifunctional linker or a homobifunctional linker, and wherein the linker comprises a maleimide group, a dipeptide moiety, a benzoic acid group or derivatives thereof, a C.sub.1-C.sub.6 alkyl group, or a combination thereof.
15. The method of claim 10, wherein the PMO conjugate has an average of PMO molecule to antibody ratio (DAR) of 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 8:1.
16. The method of claim 10, wherein the PMO conjugate has an average DAR in the range of 3.5-4.5 or 7.5-8.5.
17. The method of claim 10, wherein the PMO conjugate has an average DAR of 4 or 8.
18. The method of claim 10, wherein the PMO conjugate is administered parenterally.
19. The method of claim 10, wherein the truncated dystrophin protein modulates muscular dystrophy.
20. The method of claim 19, wherein the muscular dystrophy is Duchenne muscular dystrophy or Becker muscular dystrophy.
21. A method of inducing exon 44 skipping in a targeted pre-mRNA transcript of DMD gene, comprising: a) contacting a muscle cell with a phosphorodiamidate morpholino oligonucleotide (PMO)-antibody conjugate, wherein the PMO-antibody conjugate comprises an anti-transferrin receptor antibody or antigen binding fragment thereof, and a PMO molecule consisting of a sequence selected from the group consisting of SEQ ID NOs: 116-119; wherein the PMO molecule induces exon 44 skipping in the targeted pre-mRNA transcript, and wherein the PMO-antibody conjugate is preferentially delivered into the muscle cell; b) hybridizing the PMO molecule to the targeted pre-mRNA transcript to induce exon 44 skipping in the targeted pre-mRNA transcript; and c) translating an mRNA transcript produced from the targeted pre-mRNA transcript processed in step b) in the muscle cell to generate a truncated dystrophin protein.
19/329,403
12,071,621
SEQ ID NO: 118
cgccgccatttctcaacagatctgtc
cgccgccatttctcaacagatctgtc
SEQ ID NO: 17
YTFTNYWMH
YTFTNYWMH
SEQ ID NO: 20
EINPINGRSNYAEKFQG
EINPINGRSNYAEKFQG
SEQ ID NO: 19
GTRAMHY
GTRAMHY
SEQ ID NO: 22
RTSENIYNNLA
RTSENIYNNLA
SEQ ID NO: 23
AATNLAD
AATNLAD
SEQ ID NO: 24
QHFWGTPLT
QHFWGTPLT
SEQ ID NO: 34
SEQ ID NO: 34
SEQ ID NO: 30
SEQ ID NO: 30
SEQ ID NO: 63
SEQ ID NO: 63
SEQ ID NO: 48
SEQ ID NO: 48
A person of skill in the art, reading the claims of the ‘621 patent, would look to the patent and follow the ‘621 patent’s express instruction on how to make the product within the patent, e.g., Example 7, thereby arriving at the claimed method of the examined claims.
Conclusion
No claim is allowed.
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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