Therapeutic protein and vaccine stabilisation technology with global reach across the pharmaceutical industry
Submitting Institutions
University of Strathclyde,
University of GlasgowUnit of Assessment
ChemistrySummary Impact Type
TechnologicalResearch Subject Area(s)
Chemical Sciences: Macromolecular and Materials Chemistry
Biological Sciences: Biochemistry and Cell Biology
Medical and Health Sciences: Pharmacology and Pharmaceutical Sciences
Summary of the impact
A novel self-assembly process, developed at WestCHEM was shown to provide
a step-change for
stabilising proteins as dry powders. The spin-out company, XstalBio, was
created in 2004 and
licensed the patented technology with the aim of developing it for
delivery and formulation of
therapeutic biomolecules and vaccines. Over the period 2008-2012, eight
leading international
pharmaceutical and animal health companies paid XstalBio over £2.2M for
access to its IP portfolio
and to undertake evaluation studies with candidate biomedicines and
vaccines. XstalBio employed
8 highly skilled research scientists over this period and 4 further patent
families were generated.
Boehringer Ingelheim licensed the technology for application to its
therapeutic biomolecules and in
collaboration with XstalBio built a dedicated €5M pilot plant for
manufacture of inhalable dry
powders.
Underpinning research
Context
Medicines based on biomolecules, including vaccines, are currently the
major engines of growth in
the pharmaceutical industry, with sales predicted to increase from ~$130bn
in 2012 to ~$280bn in
2022. The in vivo activity of biomolecules such as enzymes,
monoclonal antibodies, and vaccines
is determined by the tertiary structure, i.e., the
three-dimensional conformation. This means that
biologic drugs are much less stable and therefore harder to formulate and
administer than
traditional small molecule drugs.
Research by the research groups of BD Moore (Senior Lecturer and Reader,
WestCHEM), PJ
Halling (Professor, WestCHEM) and MC Parker (Lecturer, WestCHEM) aimed to
facilitate the
transfer of active enzymes from aqueous to organic media to harness their
catalytic power in
organic synthesis. A key observation was that dehydration processes other
than lyophilisation
resulted in much better retention of enzyme catalytic activity and
excellent preservation of protein
tertiary structure in the dry state (1). The wider importance of this
finding was recognised and the
innovative stabilisation technology was translated into commercial
formulation of therapeutic
biomolecules and vaccines.
Key Findings
The pathway of transferring an enzyme from aqueous solution into
non-aqueous media is found to
be critical in maintaining catalytic activity and surprisingly,
conventional protein drying methods
such as lyophilisation give very poor results. Moore and Parker discovered
and patented ("Rapid
Dehydration of Proteins", WO0069887) a much more successful
process based on co-precipitation
of enzymes with an inorganic salt by addition of an aqueous mixture to an
excess of water-miscible
solvent (2). This method was designed to rapidly dehydrate the enzyme, as
the water dissolved
into the solvent, but the additional bonus was that it also resulted in
spontaneous formation of
microcrystals of the salt coated with protein. Self-assembly of these
protein-coated microcrystals
(PCMCs) was found to be applicable to all enzyme classes tested and the
process coupled with
control of protonation state (3) are recognised as benchmark methods for
preparing biocatalysts for
use in non-aqueous media. The high catalytic activity of enzymes on dry
PCMC particles was
postulated to be because an unusually high proportion of protein molecules
retained a native
tertiary conformation in the dry state. This was subsequently proven to be
the case by solid-state
circular dichroism spectroscopy and active site-titrations in dry solvent.
The excellent retention of protein native structure in PCMCs was
recognised to have applications
beyond biocatalysis and applied research at WestCHEM explored their
potential use as a platform
for delivery of therapeutic biomolecules by inhalation (4). This research
identified a novel method
for forming free-flowing dry powders by supercritical fluid carbon dioxide
extraction of suspensions
of PCMC in solvent. The formulations and process were patented
("Pharmaceutical Composition",
WO2004062560) in conjunction with an alternate continuous flow
process for precipitating PCMC
particles ("Process for Preparing Microcrystals",
WO2006010921). A range of physiologically
acceptable water-soluble crystals, including amino-acids and sugars, were
investigated with the
aim of preparing PCMC particles in the size-range 3-5 µm suitable for
delivery to the lung. Dry
powders of model proteins with promising aerodynamic properties were
identified and found to be
unusually stable to high temperature and humidity meaning they could be
stored without requiring
refrigeration or an inert atmosphere. These properties are highly
desirable for formulation and
delivery of biologic drugs and vaccines (5).
The spin-out of XstalBio in 2004 provided access to much more
aggregation-sensitive proteins
such as humanised monoclonal antibodies supplied by commercial partners,
and stimulated further
improvements to the technology. In research directed by Moore,
zwitterionic additives were
identified which disrupted aggregation of precipitated protein on exposure
to polar solvents and
enabled them to be incorporated onto the outer surface of the particles in
a native quaternary state
(6). These additives were patented ("Precipitation Stabilising
Compositions", WO2008132439) and
the technology has been successfully applied to the formulation of over
ten therapeutic monoclonal
antibodies in development by pharmaceutical companies.
It was also shown that a double decomposition process could be carried
out in a polar organic
solvent between (i) phosphate salts that have been co-immobilised with
biomolecules on the
crystal surface and (ii) calcium chloride dissolved in the solvent. The
resultant outer shell of
sparingly aqueous-soluble calcium phosphate can be used to tune the rate
at which biomolecules
are released back into aqueous solution. Patented in 2008 ("Slow Release
Compositions",
WO2009077732) these findings have led to the development of vaccines in
which both antigens
and toll-like receptor agonists are co-immobilised on slow-release
particles, resulting in enhanced
innate and adaptive immune responses. These are being exploited in
temperature stable vaccines
for treatment of helminths in livestock and in vivo trials are
on-going in collaboration with the
Moredun Research Institute. All of the patents filed have been licensed to
XstalBio Ltd.
Key Researchers
Barry D Moore, (employed at WestCHEM from January 1991, Senior Lecturer
from April 1997 and
Reader from June 2002); Marie Claire Parker, (employed as Lecturer at
WestCHEM from 1997-2005,
and currently Honorary Research Fellow); Peter Halling, (employed at
WestCHEM since
August 1983, Professor since August 1990).
References to the research
References 1-3 (highly cited primary papers) best exemplify the quality
of the body of research.
[1] Practical route to high activity enzyme preparations for synthesis in
organic media. Partridge,
J.; Halling, P. J.; Moore, B. D., Chem. Commun., 1998,
841-842; DOI: 10.1039/A800408K
[2] Enzyme-coated micro-crystals: a 1-step method for high-activity
biocatalyst preparation.
Kreiner, M.; Moore, B. D.; Parker, M. C., Chem. Commun., 2001,
1096-1097; DOI:
10.1039/B100722J
[3] Control of enzyme activity in organic media by solid-state acid-base
buffers. Zacharis, E;
Moore, B.D.; Halling, P. J., J. Am. Chem. Soc., 1997, 119,
12396-12397; DOI:
10.1021/ja972635c
[4] Ex vivo perfusion bioassay: an excellent technique to measure the
bioactivity of inhalable
insulin coated microcrystals. Ross A. C.; Steve H. N.; Partridge J.; Moore
B. D.; Flores M. V.;
Parker M. C.; Brown A. J.; Hillier C.; Coleman J., Abstracts of the
AAPS 2002;
http://abstracts.aaps.org/published/ContentInfo.aspx?conID=32975
[5] Formulation of the adenylate cyclase toxin of Bordetella pertussis as
protein-coated
microcrystals. Khosravani A.; Parker M. C.; Parton R.; Coote J., Vaccine.
2007, 25, 4361-4367;
DOI: 10.1016/j.vaccine.2007.03.035
Details of the impact
From research to impact
XstalBio Ltd was formed as a spin-out company in 2004 to license,
commercialise, and extend the
intellectual property associated with the protein and vaccine
stabilisation technology. XstalBio's
technology adds therapeutic value, accelerates the development, and
extends the life-cycles of
protein-based drugs as well as enabling new product opportunities. The
company targets the
$50bn currently spent per annum by the pharmaceutical industry on
biopharmaceutical product
development. XstalBio markets its expertise and technology to these pharma
companies with the
aim of enabling development of improved formulation and delivery methods
for new candidate
biologic drugs and vaccines.
In an internationally competitive market, XstalBio has succeeded in
selling contracts for access to
its Intellectual Property Portfolio for over 8 years and has continuously
developed the patented
technology to meet the new challenges facing the biopharmaceutical
industry. These contracts,
worth £2.2M over the period 2008-2012, are underpinned by option license
agreements taken out
on the WestCHEM owned patents. An early client was Boehringer Ingelheim
who subsequently
licensed the stabilisation technology from XstalBio, primarily for
delivery of biologics by inhalation.
As part of this license agreement the two companies co-developed and built
a dedicated GMP
compliant pilot plant for inhalation dry powders, which was commissioned
in Biberach, Germany in
2008 at cost of €5M.
Development programmes have also been carried out with, or are on-going
with, other major
pharmaceutical companies including [text removed for publication] with
most contracts being with
biologic drug development groups from outside the UK. The specific details
of these are subject to
stringent commercial confidentiality agreements but in general the role of
XstalBio is to accelerate
development, enhance therapeutic value, or manage the life-cycle of the
biopharmaceutical by
providing innovative dry powder formulations which improve the overall
product profile.
Type of Impact
An important impact of the original research has been economic with the
launch and growth of a
sustainable UK based SME with global reach in the pharmaceutical industry.
In the period 2008-2012,
the company made international sales of its Intellectual Property of £2.2M
and employed 8
full-time research and development scientists at BSc and PhD level. The
company has also
supported and trained 3 PhD CASE studentships based at WestCHEM and the
London School of
Pharmacy.
Vaccines and medicines based on biomolecules will be major engines of
growth in the
pharmaceutical industry over the coming decades. The significant payments
by international
companies to XstalBio for access to its IP portfolio demonstrate that its
biologic formulation and
drug delivery technologies lie at the commercial cutting-edge and are
impacting on the direction
taken by top ten pharmaceutical companies in developing the next
generation of biologic
medicines.
Unmet product and process needs that XstalBio Technology is being used to
address include:
- Delivery by inhalation with a dry powder inhaler requires sensitive
biologics to be processed
into particles in the size-range of 3-5 microns with no loss of tertiary
structure or bioactivity.
Techniques such as milling cannot be used and spray-drying produces
extremely moisture-sensitive
powders which require expensive protective packaging. The impact of
XstalBio's
PCMC technology in this field was recognised by Boehringer Ingelheim and
resulted in a
licensing agreement and significant investment in a GMP compliant
manufacturing facility
(estimated €5M),
- Shipping and storage of biomolecules without the need for continuous
refrigeration is a major
goal for organisations and companies that intend to supply diagnostics,
biological medicines
and vaccines in challenging environments such as remote regions in the
developing world.
XstalBio PCMC dry powders remain stable at high temperatures and in
humid conditions and
therefore offer considerable cost savings to its partners. Boehringer
Ingelheim has noted these
more efficient and cost effective manufacturing opportunities compared
with conventional
methods:
"Particularly with regard to cost-effectiveness, precipitation methods
are attractive. An
interesting method is the protein-coated microcrystals (PCMC) technology
that stabilises
biomolecules on crystalline surfaces by co-precipitation during rapid
solvent exchange"
- Administration of therapeutic proteins at very high concentration by
subcutaneous delivery
would provide an alternative to lengthy infusions, reducing resource use
(time in clinic) and
improving quality of life for patients with chronic conditions. As was
demonstrated with a Pfizer
candidate human monoclonal antibody, the patented stabilising additives
prevent aggregation
and conserve bioactivity. Because the dry mAb powders can also be
rapidly reconstituted to
very high protein concentration (>200mg/ml) they are providing
industry with a radical
alternative to concentrating by TFF which becomes very difficult at high
viscosities.
- Tuneable drug delivery kinetics: Slowing the release of therapeutic
proteins without covalent
modification is desirable. Use of a calcium phosphate shell allows
release of proteins from
PCMCs to be tuned from hours to days.
The collaboration with Boehringer Ingelheim illustrates how the XstalBio
technology has had an
impact on a client's R&D strategy and investment. Boehringer Ingelheim
has stated publicly that
"Boehringer Ingelheim believes that the collaboration with XstalBio
will provide improved methods
for the formulation and delivery of biomolecules. The technology
provides a highly differentiated
method for preparing biomolecules as stable, solid formulations, with
the particles capable of being
engineered for delivery in a range of formulations via various routes of
administration. Both
partners will jointly develop and scale-up the PCMC technology for GMP
manufacturing at Pilot
scale."
The publication of the 6 filed and licensed patent families impacts on
the overall knowledge base
and direction of the pharmaceutical industry. This is evidenced by the
presence of 11 patent filings
from other companies, including BASF, Novo Nordisk, Boheringher Ingelheim,
Lek, Taisho
Pharmaceutical Co. Ltd and Dominó — Indústrias Cerâmicas that reference
the technology and/or
patents.
Sources to corroborate the impact
1. The Non-Executive Chairman of XstalBio will corroborate the company's
client base and that
XstalBio's technology is being applied to solving a range of
biopharmaceutical product
challenges for those clients.
2. www.xstalbio.com/technology & bioresearchcentral.pharmaloco.com/company/XstalBio/index.html
will corroborate the claims that XstalBio's technology is being applied to
solving a range of
biopharmaceutical product challenges for its clients.
3.
http://formulation.org.uk/index.php?option=com_content&view=article&id=279&Itemid=223
will corroborate the claim that collaboration between XstalBio and Pfizer
has improved stability of
human monoclonal antibody formulations.
4.
http://www.boehringer-ingelheim.ca/en/news/press_releases/2006/16_may_2006.html
will corro-borate
Boehringer Ingelheim's relationship with XstalBio
5. "Development of a pilot-scale manufacturing process for protein-coated
microcrystals (PCMC):
Mixing and precipitation — Part I", C. König, K. Bechtold-Peters, V. Baum,
T. Schultz-Fademrecht,
S. Bassarab, K.-J. Steffens, Eur. J. Pharm. Biopharm. 2012, 80,
490-498 (DOI:
10.1016/j.ejpb.2011.11.012) will corroborate the claim that XstalBio and
Boehringer Ingelheim
have established a pilot-scale manufacturing process which will provide
improved methods for
the formulation and delivery of biomolecules.
6. World Intellectual Property Organisation (http://patentscope.wipo.int/),
will corroborate filing of
the following patents by B.D. Moore et al. at Strathclyde:
Rapid Dehydration of Proteins WO/2000/069887; Pharmaceutical
Composition
WO/2004/062560; Process for Preparing Microcrystals
WO/2006/010921; Precipitation
Stabilising Compositions WO/2008/132439; Slow Release Compositions
WO/2009/077732;
Method for preparing amorphous precipitated protein particles WO/2013/093524