Patent: 9,365,645

Summary for Patent: 9,365,645

Title:	Methods for controlling the galactosylation profile of recombinantly-expressed proteins
Abstract:	The present invention relates to methods for modulating the glycosylation profile of recombinantly-expressed proteins. In particular, the present invention relates to methods of controlling the galactosylation profile of recombinantly-expressed proteins by supplementing production medium, e.g., a hydrolysate-based or a chemically defined medium, with manganese and/or D-galactose.
Inventor(s):	Bengea; Cornelia (Auburn, MA), Rives; Lisa M. (Natick, MA), Hossler; Patrick (Westborough, MA)
Assignee:	AbbVie, Inc. (North Chicago, IL)
Application Number:	15/014,694
Patent Claims:	see list of patent claims

Executive summary: US Patent 9,365,645 is a glycoengineering-and-formulation patent family centered on adalimumab compositions with controlled N-glycan distribution, specifically limiting agalactosyl fucosylated biantennary glycans (sum NGA2F + NGA2F-GlcNAc) and tuning galactose-containing fucosylated biantennary fractions (sum NA1F + NA2F). Independent claim scope is narrow in that it is defined by measured glyco-composition percentages rather than by a specific amino-acid change or a specific antibody variant sequence. Practical validity and enforceability hinge on (i) whether the claimed glycan-spec definitions are sufficiently supported and definite under US standards, (ii) whether prior art established commercially relevant control of those exact glycan populations for adalimumab, and (iii) whether infringement can be proven with robust glycoanalytical methodology on accused lots. The manufacturing-dependent claims (manganese and galactose supplementation in the culture media; CHO/NSO; defined dose ranges) create a second infringement pathway but increase the evidentiary and freedom-to-operate (FTO) burden for generic/biosimilar makers because they must replicate the precise process conditions and still hit the glyco distribution thresholds.

US Patent 9,365,645 claims explained: What antibody glycan fractions are protected for adalimumab?

Short answer: The claims protect adalimumab (or an antibody with adalimumab heavy/light variable domains) where the N-linked glycan profile contains less than 70% of a specific agalactosyl, fucosylated biantennary group (sum NGA2F + NGA2F-GlcNAc), with optional narrower ranges that pin down 49–69%, 55–65%, or 26–40% / 35–44% / 30–40% galactose-containing fucosylated biantennary fractions (sum NA1F + NA2F).

Which glyco-specs define infringement under claim 1?

Claim 1 is the core composition claim. It sets two mutually supporting control concepts:

1. Agala-fucosylated biantennary limitation (low “Agal”):
   less than 70% of total N-linked oligosaccharides are of the agalactosyl fucosylated biantennary form defined as sum NGA2F + NGA2F-GlcNAc.

2. Binary structure definition by group sums (not individual peaks):
   The glyco analytics are framed as sums of defined species, implying the patent’s claim boundaries track aggregate class fractions rather than a single peak intensity.

Claims 3–7 progressively narrow the agalactosyl fucosylated biantennary fraction and/or specify galactose-containing fucosylated biantennary fractions:

• Claim 3: 49–69% (agalactosyl fucosylated biantennary sum)
• Claim 4: 55–65% (agalactosyl fucosylated biantennary sum)
• Claim 5: 26–40% (galactose-containing fucosylated biantennary sum NA1F + NA2F)
• Claim 6: 35–44%
• Claim 7: 30–40%

Is claim scope limited to “adalimumab” or broader?

• Claim 2, 16: explicitly cover adalimumab where those glycan fractions are within the defined bounds.
• Claim 1: covers an antibody comprising adalimumab heavy and light chain variable domains, even if framed outside the word “adalimumab.” That matters for enforceability against engineered “adalimumab-like” antibodies with the same variable domains but differing constant regions, if those are within the claim’s wording.

How do the two “sum” definitions interact in practice?

The claims are structured as two axes of glyco control:

• Reduce agalactosyl fucosylated biantennary fraction (NGA2F + NGA2F-GlcNAc).
• Establish a bounded fraction of galactose-containing fucosylated biantennary (NA1F + NA2F).

For infringement, an accused product must meet the specific percentage gates that correspond to the claim being asserted. Because the claims include both “less than 70%” and narrower percentage bands, a product can be outside claim 1 but still fall into a dependent-range claim if the measured values align. The dependency drafting creates a ladder effect that can be used strategically in litigation: if the accused product is not at the strictest range, the patentee can still attempt the broader “<70%” claim.

How would courts interpret “total N-linked oligosaccharides”?

The phrase “total N-linked oligosaccharides present on said antibody” points to a normalization denominator. That normalization is a frequent litigation pressure point:

• If the accused party uses different digestion, labeling, chromatographic separation, or peak integration conventions, the computed percentages can shift.
• The claim language’s reliance on group sums increases the likelihood of disputes about which measured glycoforms map to NGA2F, NGA2F-GlcNAc, NA1F, and NA2F under the accused lab’s method.

What manufacturing-process claims are included in US 9,365,645 and how do they expand infringement risk?

Short answer: Claims 8–11 and 22–25 add process conditions: culturing a mammalian cell in media supplemented with manganese and galactose (with defined concentration ranges), using either hydrolysate-based or chemically defined media, and employing CHO or NSO cells.

Process conditions in the claims

The manufacturing-dependent claims add elements that can be litigated via discovery of:

• batch records,
• raw material specifications (manganese and galactose forms and grades),
• media recipes and supplements,
• culture conditions logs,
• and release testing documentation connecting culture inputs to glyco outcomes.

Key claim elements:

Manganese + galactose supplementation

• Claim 8/22: media supplemented with a manganese supplement and a galactose supplement.
• Claim 9/23:
  • manganese: 0.2–100 μM
  • galactose: 1–100 mM
• Claims 10/24: media type includes hydrolysate-based media or chemically defined media.
• Claims 11/25: mammalian cell is CHO or NSO.

Why these process claims matter even if glyco-specs are met

A biosimilar maker could design a process that yields the glycan profile without using the exact manganese/galactose window. In that scenario:

• A product might still fall within the composition-by-glyco-spec claims (claims 1/2 or 16), even if it avoids the process.
• Conversely, even if a maker’s glyco profile is slightly outside the rigid thresholds, the process claims can still be used if plaintiffs can show that the accused process predictably yields within-range material at commercial scale. Practice varies by jurisdiction, but the process claims often increase litigation leverage.

FTO implication for alternative manufacturing

If competitors use different glyco-manipulation levers (e.g., different enzyme knockdowns, glycosyltransferase expression tuning, or alternative supplements), they may reduce risk of hitting the process claim elements, but they cannot ignore the composition claims defined by the final glyco distribution.

What pharmaceutical formulation claims does US 9,365,645 cover beyond glycosylation?

Short answer: The patent also claims parenteral pharmaceutical compositions comprising the glyco-defined adalimumab composition plus a pharmaceutically acceptable carrier, with additional limitations around polyalcohol and excipient exemplars (mannitol, sodium chloride, sodium phosphate, polysorbate 80).

Formulation claim structure

• Claim 12/26: pharmaceutical composition comprising the glyco-defined composition + pharmaceutically acceptable carrier.
• Claim 13/27: carrier comprises a polyalcohol.
• Claim 14/28: carrier comprises one or more of: mannitol, sodium chloride, sodium phosphate, polysorbate 80.
• Claim 29: further comprises polysorbate 80 (dependent layering).
• Claim 15/30: suitable for parenteral administration.

Criticality for infringement

These formulation claims can be easier to satisfy for infringement because typical adalimumab formulations commonly include excipients like polysorbate 80 and salts, depending on the marketed product and concentration. The novelty risk is not the formulation excipients themselves; it is the combination of excipient carrier + the glyco-defined antibody. The formulation layer can become the hook for asserting infringement against lots that meet both the glyco specification and the standard parenteral excipient strategy.

How strong is US 9,365,645 for validity against glycoengineering prior art?

Short answer: The strength of the estate depends on whether pre-filing art already disclosed adalimumab glyco-population control that matches the claimed percentage gates, and whether culture supplementation with manganese and galactose at the claimed ranges was known to drive the same glyco fractions for IgG1 Fc N-glycans. The more generic the prior art on “improved galactosylation” without matching these exact group-sum thresholds, the more the patent’s enforceability leans on the specificity of the numeric ranges and the particular species mapping.

Where prior art tends to overlap

Glycoengineering literature and platform patents commonly disclose:

• raising galactosylation (GA) fractions,
• reducing agalactosylation (G0/agalactosyl) and/or fucosylated agalactosyl species,
• and using culture media supplements that modulate glycosylation capacity.

The vulnerability point for US 9,365,645 is that broad disclosure that “supplements such as Mn and galactose change Fc glycosylation” may be used to argue obviousness if it is also linked to the kind of measurable distributions the patent claims.

Where the patent gains distinctiveness

The distinctiveness comes from:

• the specific agalactosyl, fucosylated biantennary sum (NGA2F + NGA2F-GlcNAc) capped at defined thresholds,
• and the dependent-range ladder that tightly bounds those percentages,
• plus the inclusion of defined manganese and galactose concentration ranges.

If pre-filing art does not disclose the same numeric ranges on the same glycoform mapping scheme, validity arguments become fact-intensive.

Definiteness and measurement-method risk

Even if prior art is close, validity and infringement hinge on whether the glyco-spec definitions are sufficiently definite and whether they can be reproduced. Numeric glyco fractions are inherently dependent on:

• analytical platforms (e.g., LC-MS profiling vs fluorescent labeling workflows),
• integration conventions,
• calibration standards,
• and assignment rules for NGA2F vs NGA2F-GlcNAc and NA1F vs NA2F.

Under US litigation practice, plaintiffs typically need to connect their method to the claim-defined terms. The accused infringer typically attacks mapping and denominators.

When does US Patent 9,365,645 expire and how would that affect generic or biosimilar entry timing?

Short answer: No expiration date can be computed from the claim text alone. US utility patent term generally runs from earliest non-provisional filing plus adjustments, but those facts are not provided in the prompt, so timing analysis cannot be stated accurately.

What is the Orange Book status of US 9,365,645 for adalimumab?

Short answer: Orange Book listing status cannot be determined from the prompt. Orange Book coverage depends on the patent’s publication and listing actions for a specific approved NDA product, and no product-patent listing data is included here.

How do Paragraph IV challenges map to an antibody patent focused on glycosylation?

Short answer: The prompt does not identify any FDA approval pathway for the specific challenged product, any listed patents, or any litigation filings. Without that, mapping to Paragraph IV risk is not actionable.

What biosimilar risk does US 9,365,645 create for adalimumab biosimilars?

Short answer: The patent’s glyco-spec and process supplement claims create biosimilar exposure at two levels:

1. Analytical matching risk: a biosimilar that achieves clinically acceptable “similarity” may still land inside the claimed glyco fraction windows depending on formulation and upstream process.
2. Process parameter exposure: if the biosimilar uses manganese and galactose supplementation in the defined windows with CHO/NSO, it can implicate the process claims as well.

Practical litigation leverage

Because the patent claims are anchored to quantitative glycan profiles, enforcement often turns on:

• head-to-head glyco profiling of commercial lots,
• expert comparison of glycoform assignment and fraction computation,
• and evidence that manufacturing inputs correlate with the final glyco distribution.

Which design-arounds are most plausible against US 9,365,645 based on the claim structure?

Short answer: The claims suggest design-around strategies that shift either the final glycan fractions outside the numeric bands or the process conditions outside the claimed media supplement windows.

Design-around vector 1: Move glyco profile outside the claimed thresholds

Because claim 1 caps agalactosyl fucosylated biantennary below 70%, a manufacturer can try to:

• keep NGA2F + NGA2F-GlcNAc at or above 70%, or
• keep galactose-containing fucosylated biantennary outside the NA1F + NA2F ranges tied to dependent claims.

This may conflict with quality-by-design goals, product comparability, and immunogenicity targets, but it is conceptually aligned with the claim boundary.

Design-around vector 2: Avoid the specific manganese and galactose ranges

A manufacturer could:

• omit manganese supplementation,
• use manganese outside 0.2–100 μM,
• use galactose outside 1–100 mM,
• or change cell line beyond CHO/NSO.

That would aim to defeat the process-dependent claims while leaving open the possibility of meeting glyco-specs under composition claims.

Design-around vector 3: Change downstream formulation excipients

Because formulation claims include commonly used excipients, excipient-only design-around may not be meaningful. The formulation layer is secondary; defeating it without defeating glyco-spec claims is unlikely to eliminate risk.

How does US 9,365,645 compare with broader adalimumab glycoengineering estates?

Short answer: US 9,365,645 is more specific than platform patents that generally claim “altering glycosylation” because it:

• defines glyco fractions by summed species,
• anchors to particular numeric ranges,
• and adds narrow process supplement concentration windows.

That specificity usually improves the patentee’s infringement narrative but can raise validity risk if prior art contains the same numeric glyco outcomes or if measurement definitions were already standardized.

Key takeaways

• US 9,365,645 is a glyco-spec patent: infringement is driven by quantitative N-glycan population limits on adalimumab (or adalimumab variable-domain antibodies).
• Dependent claims tighten the glyco distribution into specific numeric windows, creating multiple potential infringement “targets” across the same analytical framework.
• Process-dependent claims expand coverage by requiring manganese and galactose supplementation in defined concentration windows using CHO/NSO, alongside media-type constraints.
• Formulation claims extend the estate into standard parenteral carrier combinations (polyalcohol; exemplar excipients including polysorbate 80), but the core novelty is the glyco-defined antibody.
• Litigation strength typically depends on glyco-analytical reproducibility, species mapping (NGA2F vs NGA2F-GlcNAc; NA1F vs NA2F), and whether accused lots or upstream recipes land inside the numeric gates.

FAQs

1. What analytical methods are typically used to quantify NGA2F and NA1F fractions in IgG Fc glycan profiles for infringement?
2. Can a biosimilar that matches clinical glycosylation “similarity” still infringe numeric glyco-spec patents like US 9,365,645?
3. Do manganese and galactose supplements always correlate with reduced agalactosyl fucosylated biantennary species in CHO/NSO systems?
4. How do excipient changes (polysorbate 80, mannitol, salts) affect formulation-dependent claims for parenteral antibodies?
5. What evidence most often decides whether a defendant’s glyco-spec measurements satisfy “total N-linked oligosaccharides” denominators?

References

1. US Patent 9,365,645 (claims provided in prompt).

Details for Patent 9,365,645

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	December 31, 2002	9,365,645	2036-02-03
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	February 21, 2008	9,365,645	2036-02-03
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	April 24, 2013	9,365,645	2036-02-03
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	September 23, 2014	9,365,645	2036-02-03