Patricia Van Arnum was executive editor of Pharmaceutical Technology.

Patricia Van Arnum

The International Society for Pharmaceutical Engineering (ISPE) has released the latest volume in the Product Quality Lifecycle Implementation (PQLI) series, 

Change Management System as a Key Element of a Pharmaceutical Quality System

. The guide, which is Part 3 of the PQLI series, provides practical, real-world strategies that companies can use to implement the change-management recommendations of ICH Q10. 

The PQLI guide on change management outlines potential product life-cycle approaches to the change-management system of a pharmaceutical quality system (PQS). It provides guidance on achieving compliant and consistent change-management systems and describes how to implement compliant change-management systems within a PQS. 

 can be used in conjunction with the first two PQLI guides as part of a larger quality-by-design approach or as a stand-alone change-management manual.

Articles

New guide outlines potential product life-cycle approaches to the change-management system of a pharmaceutical quality system.

ISPE Issues Change-Management Guide

As a drug type, peptides offer certain benefits, such as specificity and potency, but they also present challenges, such as poor stability and short half-life. Recent partnerships among large pharmaceutical companies and specialized companies as well as advances from academia are seeking to resolve these problems. 

 Chad Baker/Digital Vision/Getty Images			

Peptides and proteins have the size and functionality to effectively modulate intracellular protein–protein interactions, but they often do not permeate cells and therefore are used to modulate extracellular targets such as receptors (1, 2). The majority of peptide candidates target extracellular molecules with less than 10% binding to intracellular targets, according to a recent analysis of the peptide drug pipeline by the Peptide Therapeutics Foundation (3). The most common extracellular targets were G-protein coupled receptors (GPCR), which include nearly 1000 transmembrane proteins that activate cellular response. During 2000–2008, 60% of peptides entering clinical development targeted GPCRs, and most had agonist activity (3). 

Although a small portion of total drug candidates, the number of peptide drugs entering clinical development has increased during the past several decades, according to the Peptide Therapeutics Foundation analysis, which excluded insulins (3). The study found that the average number of new peptide candidates entering clinical development in the 1970s was 1.2 per year and rose to 4.6 per year in the 1980s, 9.7 per year in the 1990s, and 16.8 per year through 2000–2008 (3). During 2000–2008, peptides entering clinical study were most frequently treatments for cancer and metabolic disorders (including diabetes and obesity), respectively, representing 18% and 17% of peptide drug development. Decreases were observed for peptides studied as therapies for treating allergies, immunological disorders and cardiovascular diseases (3). 

On a commercial level, several peptide-based therapeutics have reached blockbuster status, defined as having sales of $1 billion or more, or near blockbuster status (3). These drugs, using 2011 global sales figures from company annual financial reports, include:  					

Teva Pharmaceutical's Copaxone (glatiramer acetate), an L-glutamic acid polymer with L-alanine, L-lysine and L-tyrosine (2011 global sales of $3.6 billion)

Abbott's Lupron (leuprolide acetate), a synthetic nonapeptide analog of the naturally occurring gonadotropin-releasing hormone (GnRH or luteinizing hormone-releasing hormone [LHRH]) (2011 global sales of $810 million)

AstraZeneca's Zoladex (goserelin acetate), a decadpeptide and GnRH agonist and synthetic analog of a naturally occurring LHRH) (2011 global sales of $1.1 billion)

Novartis' Sandostatin (octreotide acetate), a cyclic octapeptide with pharmacologic actions mimicking those of the natural hormone somatostatin  (2011 global sales of $1.4 billion)

Eli Lilly/Amylin Pharmaceuticals' Byetta (exenatide), a 39-amino acid peptide amide (2011 global sales, Eli Lilly, $423 million, Amylin, $518 million)

Forteo (teriparatide recombinant), which contains recombinant human parathyroid hormone (1–34), is also called rhPTH (1–34). It has an identical sequence to the 34 

-terminal amino acids (the biologically active region) of the 84-amino acid human parathyroid hormone (2011 global sales of $951 million) (4–9).

				  One important collaboration in peptide drug development is between the biopharmaceutical company Aileron Therapeutics and Roche. In November 2011, Aileron expanded its collaboration with Roche for the discovery, development, and commercialization of stapled-peptide drugs. The potential $1.1-billion drug-development collaboration, launched in August 2010, encompasses up to five programs with the initial two programs focused on oncology and then a third program launched late last focused on inflammatory diseases. 

Academic and industry collaboration in pharmaceutical sciences			

Stapled peptides use peptide-stabilization technology to enhance potency and cell permeability of a drug to address pharmacological limitations of small molecules and existing biologics in intracellular protein–protein interactions. Although small molecules are able to penetrate cells, the large binding surfaces for intracellular protein–protein interactions often make small-molecule modulators ineffective. Although peptides and proteins have the size and functionality to effectively modulate intracellular protein–protein interactions, they often do not permeate cells and therefore are used to modulate extracellular targets, such as receptors (1, 10). Stapled peptides seek to resolve those problems. Because many undruggable therapeutic targets include those protein–protein interactions in which alpha-helices are required in lock-and-key-type mechanisms, an approach is to design alpha-helical peptides that have structural and functional properties that enable them to penetrate into the cell, bind to the therapeutic target, and modulate the biological pathway (1, 10). Aileron stabilizes peptides by "stapling" them with hydrocarbon bonds into an alpha-helix. Once constrained in the alpha-helix structure, the peptides are protected from degradation by proteases. The stabilized alpha-helical peptides can penetrate cells by energy-dependent active transport and typically have a higher affinity to large protein surfaces (1, 2, 10). 

Researchers at the New York Structural Biology Center recently reported on high-resolution solution nuclear magnetic resonance techniques with dynamic light-scattering to characterize a family of hydrocarbon-stapled peptides with known inhibitory activity against HIV-1 capsid assembly to evaluate the various factors that modulate activity. The researchers reported that helical peptides share a common binding motif but differ in charge, the length, and position of the staple. The research showed that the peptides share a propensity to self-associate into organized polymeric structures mediated predominantly by hydrophobic interactions between the olefinic chain and the aromatic side-chains from the peptide. The researchers also detailed the structural significance of the length and position of the staple and of olefinic bond isomerization in stabilizing the helical conformation of the peptides as potential factors influencing polymerization (11).  

. In April 2012, researchers at Carnegie Mellon University reported on their manufacturing method for a synthetic form of a cyclic peptide. Macrocyclic peptides with multiple disulfide cross-linkages, such as those produced by plants and those found in nonhuman primates, hold potential as drugs because of their broad biological activities and high chemical, thermal, and enzymatic stability (12). Because of  their intricate spatial arrangement and elaborate interstrand cross-linkages, some macrocyclic peptides are difficult to prepare in large quantities and high purity because of  the nonselective nature of disulfide-bond formation (12).  

In the current study, the Carnegie Mellon researchers focused on RTD-1, a cyclic peptide held together by three disulfide bonds. RTD-1 has a broad range of antibacterial, antifungal and antiviral capabilities and has been shown to inhibit HIV from entering cells. The researchers created a mimic of RTD-1. While the outside of the mimic peptide maintained the same amino acids as the original, the researchers replaced the disulfide bonds at its center with noncovalent Watson–Crick hydrogen bonds without significantly affecting the biological activity of the peptide. The researchers say the work provides a general strategy for engineering conformationally rigid, cyclic peptides without the need for disulfide-bond reinforcement (12). 

They tested the efficacy of the mimic RTD-1 by mixing the peptide with 

—bacteria that RTD-1 typically protects against. 

The mimic proved to be effective in killing each of the types of bacteria tested by the researchers, which included both gram-positive and gram-negative bacterial strains. Furthermore, the mimic peptide worked by binding to the bacteria's cell membrane, not its DNA or RNA, decreasing the probability that the bacteria could develop resistance to the peptide, according to an Apr. 13, 2012, Carnegie Mellon University press release. The researchers plan to see if the mimic RTD-1 is effective against other types of pathogens, including antibiotic-resistant bacteria. They also plan to apply their method to manufacture mimics of other cyclic peptides, according to the university release. 

Other approaches. In 2011, the biopharmaceutical company Sutro Biopharma formed a multiyear collaboration with Pfizer for the research, development, and commercialization of novel peptide-based therapeutics. The partnership gives Pfizer access to peptides that have been difficult to produce using conventional technologies using  a biochemical protein synthesis technology platform developed by Sutro, In 2008, Pfizer acquired CovX, a biopharmaceutical company with a technology platform that links therapeutic peptides to an antibody scaffold. The peptide targets the disease while the antibody scaffold allows the peptide to remain in the body long enough to achieve therapeutic benefit. The technology thereby allows for better half-life extension and bioavailability to support optimal dosing regimens for peptide therapeutics.  

 485 Route One South, Bldg F, First Floor, Iselin, NJ 08830 tel. 732.346.3072, 

Development Trends for Peptide Therapeutics Report

 (Feb. 27, 2009), Drugs@FDA, accessed May 15, 2012. 

 (Mar. 28, 2012), Drugs@FDA, accessed May 15, 2012. 

 (Jan. 14, 2011), Drugs@FDA, accessed May 15, 2012. 

 (Mar. 23, 2012), Drugs@FDA, accessed May 15, 2012. 

 (Oct. 19, 2011), Drugs@FDA, accessed May 15, 2012. 

 (July 26, 2009), Drugs@FDA, accessed May 15, 2012. 

Peptides and related technologies to are starting to improve production.

Peptides Gain Traction in Drug Development

As a drug type, peptides offer certain benefits, such as specificity and potency, but they also present challenges, such as poor stability and short halflife. Recent partnerships among large pharmaceutical companies and specialised companies, as well as advances from academia, are seeking to resolve these problems.  

Peptides and proteins have the size and functionality to effectively modulate intracellular protein–protein interactions, but they often do not permeate cells and, therefore, are used to modulate extracellular targets such as receptors (1, 2). According to a recent analysis of the peptide drug pipeline by the Peptide Therapeutics Foundation (3), the majority of peptide candidates target extracellular molecules with less than 10% binding to intracellular targets. The most common extracellular targets were G-protein coupled receptors (GPCR), which include nearly 1000 transmembrane proteins that activate cellular response. During 2000–2008, 60% of peptides entering clinical development targeted GPCRs and most had agonist activity (3). 

Although making up only a small portion of total drug candidates, the number of peptide drugs entering clinical development has increased during the past several decades. The Peptide Therapeutics Foundation analysis, which excluded insulins, found that the average number of new peptide candidates entering clinical development in the 1970s was 1.2 per year, rising to 4.6 per year in the 1980s, 9.7 per year in the 1990s and 16.8 per year through 2000–2008 (3). During 2000–2008, peptides entering clinical study were most frequently treatments for cancer and metabolic disorders (including diabetes and obesity), respectively, representing 18% and 17% of peptide drug development. Decreases were observed for peptides studied as therapies for treating allergies, immunological disorders and cardiovascular diseases. 

On a commercial level, several peptide-based therapeutics have reached blockbuster status, defined as having sales of $1 billion or more, or near blockbuster status (3). These drugs, using 2011 global sales figures from company annual financial reports, include:   

Teva Pharmaceutical's Copaxone (glatiramer acetate), an L-glutamic acid polymer with L-alanine, L-lysine and L-tyrosine; (2011 global sales of $3.6 billion).

Abbott's Lupron (leuprolide acetate), a synthetic nonapeptide analogue of the naturally occurring gonadotropin-releasing hormone (GnRH or luteinising hormone-releasing hormone [LHRH]); (2011 global sales of $810 million).

AstraZeneca's Zoladex (goserelin acetate), a decadpeptide and GnRH agonist and synthetic analogue of a naturally occurring LHRH); (2011 global sales of $1.1 billion),

Novartis' Sandostatin (octreotide acetate), a cyclic octapeptide with pharmacologic actions mimicking those of the natural hormone somatostatin; (2011 global sales of $1.4 billion).

Eli Lilly/Amylin Pharmaceuticals' Byetta (exenatide), a 39-amino acid peptide amide; (2011 global sales, Eli Lilly, $423 million, Amylin, $518 million).

Forteo (teriparatide recombinant), which contains recombinant human parathyroid hormone (1-34), is also called rhPTH (1-34). It has an identical sequence to the 34 

terminal amino acid (the biologically active region) of the 84-amino acid human parathyroid hormone; (2011 global sales of $951 million) (4–9).

. One important collaboration in peptide drug development is between the biopharmaceutical company Aileron Therapeutics and Roche. In November 2011, Aileron expanded its collaboration with Roche for the discovery, development and commercialisation of stapledpeptide drugs. The potential $1.1billion drug development collaboration, launched in August 2010, encompasses up to five programmes with the initial two programmes focusing on oncology, and a third programme being launched to focus on inflammatory diseases. 

Stapled peptides use peptide-stabilisation technology to enhance potency and cell permeability of a drug to address pharmacological limitations of small molecules and existing biologics in intracellular protein–protein interactions. Although small molecules can penetrate cells, the large binding surfaces for intracellular protein–protein interactions often make small-molecule modulators ineffective. As cited previously, peptides cannot permeate cells so act extracellularly (1, 10). Stapled peptides seek to resolve this problem. Because many undruggable therapeutic targets include those protein–protein interactions, in which alpha-helices are required in lock-and-key-type mechanisms, an approach is to design alpha-helical peptides that have structural and functional properties that enable them to penetrate into the cell, bind to the therapeutic target and modulate the biological pathway (1, 10). Aileron stabilises peptides by "stapling" them with hydrocarbon bonds into an alpha-helix. Once constrained in the alpha-helix structure, the peptides are protected from degradation by proteases. The stabilised alphahelical peptides can penetrate cells by energy-dependent active transport and typically have a higher affinity to large protein surfaces (1, 2, 10). 

Publicâprivate partnerships in biomanufacturing  

Researchers at the New York Structural Biology Centre recently reported on high-resolution solution nuclear magnetic resonance techniques with dynamic lightscattering to characterise a family of hydrocarbon-stapled peptides with known inhibitory activity against HIV-1 capsid assembly to evaluate the various factors that modulate activity. The researchers reported that helical peptides share a common binding motif, but differ in charge, the length and position of the staple. The research showed that the peptides share a propensity to self associate into organised polymeric structures mediated predominantly by hydrophobic interactions between the olefinic chain and the aromatic side-chains from the peptide. The researchers also detailed the structural significance of the length and position of the staple and of olefinic bond isomerisation in stabilising the helical conformation of the peptides as potential factors influencing polymerisation (11).  

. In April 2012, researchers at Carnegie Mellon University (Pennsylvania, USA) reported on their manufacturing method for producing a synthetic form of a cyclic peptide. Macrocyclic peptides with multiple disulphide crosslinkages, such as those produced by plants and those found in nonhuman primates, hold potential as drugs because of their broad biological activities and high chemical, thermal and enzymatic stability (12). Because of their intricate spatial arrangement and elaborate interstrand crosslinkages, some macrocyclic peptides are difficult to prepare in large quantities and high purity because of the nonselective nature of disulphide-bond formation (12).  

In the current study, the Carnegie Mellon researchers focused on RTD-1, a cyclic peptide held together by three disulphide bonds. RTD-1 has a broad range of antibacterial, antifungal and antiviral capabilities, and has been shown to inhibit HIV from entering cells. The researchers created a mimic of RTD-1. While the outside of the mimic peptide maintained the same amino acids as the original, the researchers replaced the disulphide bonds at its centre with noncovalent Watson–Crick hydrogen bonds without significantly affecting the biological activity of the peptide. The researchers say the work provides a general strategy for engineering conformationally rigid, cyclic peptides without the need for disulphide-bond reinforcement (12). 

— bacteria that RTD-1 typically protects against. The mimic proved to be effective in killing each of the types of bacteria tested by the researchers, which included both gram-positive and gram-negative bacterial strains. Furthermore, the mimic peptide worked by binding to the bacteria's cell membrane, not its DNA or RNA, decreasing the probability that the bacteria could develop resistance to the peptide. The researchers plan to see if the mimic RTD-1 is effective against other types of pathogens, including antibiotic-resistant bacteria. They also plan to apply their method to manufacture mimics of other cyclic peptides. 

. In 2011, the biopharmaceutical company Sutro Biopharma formed a multiyear collaboration with Pfizer for the research, development and commercialisation of novel peptidebased therapeutics. The partnership gives Pfizer access to peptides that have been difficult to produce using conventional technologies using  a biochemical protein synthesis technology platform developed by Sutro. In 2008, Pfizer acquired CovX, a biopharmaceutical company with a technology platform that links therapeutic peptides to an antibody scaffold. The peptide targets the disease while the antibody scaffold allows the peptide to remain in the body long enough to achieve therapeutic benefit. The technology thereby allows for better half-life extension and bioavailability to support optimal dosing regimens for peptide therapeutics. 

(Feb. 27, 2009), Drugs@FDA, accessed May 15, 2012. 

Although representing a small part of overall drug development, peptides and related technologies to improve their production are making inroads.

GlaxoSmithKline (GSK) began its previously announced tender offer to acquire all of the outstanding shares of Human Genome Sciences (HGS) for $13.00 per share in cash or $2.6 billion. GSK, which first made the unsolicited bid for HGS in mid-April, is proceeding with its offer despite having the bid rejected by HGS when it was first made.

The closing of the tender offer is subject to the terms and conditions detailed in the offer document. The tender offer and withdrawal rights are scheduled to expire on June 7, 2012, unless otherwise extended.

, HGS confirmed that it was aware of GSK’s intention to proceed with the $2.6-billion offer. The company will review the offer with its board of directors in consultation with independent financial and legal advisors. HGS says it plans to advise stockholders within 10 days of when GSK commenced the offer, which was May 10, 2012. 

The proposed $13.00-per-share price is the same price that GSK had offered in mid-April when HGS rejected the bid. At that time, HGS authorized the exploration of strategic alternatives, including, but not limited to, a potential sale of the company, and invited GSK to participate in that process. 

HGS posted a net loss of $381.1 million in 2011 on revenues of $131 million and also posted a loss in 2010 of $233 million. Last year, HGS received FDA approval for Benlysta (belimumab), a drug to treat lupus. The drug was approved by FDA in March 2011 and posted 2011 revenues of $52.3 million. In 2006, HGS and GSK entered into a codevelopment and commercialization agreement under which HGS conducted the Benlysta Phase II trials, with assistance from GSK. The companies share equally in Phase III/IV development costs, sales, and marketing expenses, and profits of any product commercialized under the agreement. GSK also has two late-stage candidates to which HGS has substantial financial rights, darapladib, in Phase III development for treating cardiovascular disease, and albiglutide, in Phase III development for treating Type II diabetes. 

 GlaxoSmithKline: A Rebuffed Suitor for the Moment

GlaxoSmithKline tenders offer of $13.00 per share in an unsolicited bid despite an earlier rejection by Human Genome Sciences.

GlaxoSmithKline Moves Forward with $2.6 Billion Offer for Human Genome Sciences

	The potential of single-cell genomics took a step forward recently with the announcement by The Broad Institute and Fluidigm Corporation of a new research center focused on developing research methods and discoveries in mammalian single-cell genomics. The Single-Cell Genomics Center is expected to act as a hub for collaboration among single-cell genomics researchers in many fields, including stem cells and cancer biology.

	Single-cell genomics holds much potential for more targeted drug discovery through a better understanding of cells and their interactions. Heterogeneity exists among cells in tissue samples and other populations, but this cellular variability is masked by averaging data across pooled cell samples. The ability to tease out single-cell genomic data has historically been limited by a lack of standardized, user-friendly methods that would allow the study of individual cellular variability at high definition, high throughput, and low cost, according to a Fluidigm 

. Advances in technology, such as microfluidic chips and high-throughput instruments, have made single-cell studies feasible by converting cellular heterogeneity from a source of background noise to a source of information to enable discoveries.

	According to Fluidigm, the company’s technology provides the capabilities required to analyze single cells, namely microfluidics and sensitivity at the nanoscale level, parallel processing of a large number of cells, and interrogation of a large number of gene targets. The center will be housed at the Broad Institute in Cambridge, Massachusetts, and will feature Fluidigm single-cell tools, protocols, and technologies. The center intends to develop novel single-cell, microfluidic approaches for gene-expression profiling, RNA/DNA sequencing ,and epigenetic analysis. The goal of these efforts is to make single-cell research accessible to the greater scientific community by developing and disseminating new workflows, reagents, bioinformatics tools, and data sets.

	It is hoped that these advances will provide the tools for better understanding the underlying causes of many diseases, including the progression of individual cancers, differential immune responses, and the maturation of stem cells. Targeted genomic information is key to understanding disease states for developing better therapies, an important ongoing need in current and future drug discovery.

The potential of single-cell genomics took a step forward recently with the announcement by The Broad Institute and Fluidigm Corporation of a new research center focused on developing research methods and discoveries in mammalian single-cell genomics.

Single-Cell Genomics Advancing Molecular Biology

Sanofi’s subsidiary Genzyme has received approval from FDA and EMA for  a second operation for a fill–finish product at its Waterford, Ireland, manufacturing plant. 

With this approval, Genzyme has nearly doubled its ability to fill–finish Myozyme and Lumizyme (alglucosidase alfa) produced at the 4000-L bioreactor scale. Genzyme also will begin the process to secure FDA and EMA approvals to fill–finish additional products in the second suite, with the long-term goal to use the Waterford site as a fill–finish platform across its portfolio of products.

Genzyme’s Waterford facility has been in operation for more than 10 years, and a $150-million expansion was completed earlier this year. The Waterford manufacturing site employs over 500 people and is considered Genzyme’s center of excellence for aseptic manufacturing.

Sanofi has received FDA and EMA approval for expansion of fill and finish operations for alglucosidase alfa.