0:35 Thank you. I also want to introduce my colleague, Dr Chris Godhart.
0:47 Chris is Director of Technical Marketing.
0:52 The talk is going to be divided into two parts.
0:55 Part 1 is a case study of green chemistry, and Part 2 is an overview of CPC Scientific, a corporate presentation.
1:07 This is a picture of our 1000-litre reactor vessel.
1:13 This is the M peptide case study.
1:22 This case study involves a peptide conjugated to a small molecule. The peptide sequence is known, but the small molecule's identity is proprietary.
1:34 We do a lot of these conjugates to either target tissue, bring payload, or introduce it through a membrane.
1:54 The main thing to note is that the small molecule is very sensitive to acid, so we can't use solvents like TFA. This led to the green approach for this peptide.
2:25 As a peptide CDMO, we often get questions about why we use solid phase synthesis instead of solution phase. The advantages of solid phase include the ability to easily wash away excess reagents and use multiple equivalents to force the reaction to completion.
2:52 The biggest advantage is that intermediates do not need to be isolated, purified, or characterized, which is crucial in GMP.
3:14 Solid phase synthesis, pioneered by Bruce Merrifield, allows for longer sequences with greater efficiencies and faster production. However, it also generates a lot of chemical waste due to the use of solvents.
3:47 We focus on mitigating the use of DMF, diethyl ether, and TFA in our process.
4:08 This is a small glass vessel used for solid phase synthesis. Typically, we use larger vessels. The solid support is usually polystyrene with an associated linker.
4:21 We bring in the Fmoc amino acid, protect it with piperidine, and perform washes using DMF, which creates a lot of waste.
4:43 The process involves putting solvents and reactants in, and waste comes out. At the end, we cleave the peptide using TFA and purify it with reverse phase chromatography and then we perform characterisation. In the case of liquid phase/solution phase this would have to be done for every single coupling but this isn’t the case with solid phase.
5:10 We aim to reduce or eliminate the use of solvents like diethyl ether, TFA, and DMF.
5:25 Most of the solvents used are for washing, not for the actual reaction. Only a very small part is composed of the amino acids that make up the product.
5:57 Diethyl ether is hazardous both environmentally and to the user. DMF and TFA are also in this hazardous category. We aim to replace it with more environmentally friendly solvents like aqueous ethanol and isopropanol.
6:49 Different kinds of resins can be used in solid phase, such as chlorotrityl resin and Wang resin. DMF is always used for both washing and as a coupling solvent.
7:02 We can introduce co-solvents like isopropanol in the washing steps, but not in the removal or coupling steps.
7:32 Isopropanol can be used in the washing steps, which constitute more than 80% of the solvents used in the whole cycle. We need DMF because it swells the resin, whereas isopropanol shrinks the resin. So instead of replacing DMF with isopropanol we will introduce it as a co-solvent.
8:07 We use various protection groups for hydroxy groups, we have hydroxy prolines, hydroxy-tyrosine’s and lysine. We typically protect the hydroxy groups with T-butyl, which require large amounts of TFA.
9:04 We need such high volumes to precipitate the peptide after cleavage and wash out scavengers. For this step you don’t need super strong acids.
9:42 For the M peptide, since the molecule is acid sensitive we cannot use TFA. For the coupling of lysine we use DDE, instead of using TFA. The main strategy is to leave all hydroxy groups unprotected. It is important to protect these groups during the cleavage. If you don’t use TFA you don’t really need those protection groups.
10:14 So in our approach we leave all hydroxy groups unprotected, using DDE to replace TBOC and reduce the need for strong acids.
10:55 We compare conventional and green methodologies, showing that the green approach improves atom economy and reduces the use of hazardous solvents. Conventional SPPS uses CTC resin or wang resin but wang resin needs a strong acid. In our case study everything is unprotected except for the lysine. When the resin is cleaved off, we can use dilute hydrazine.
11:46 Dichloromethane is used instead of DMF, as it is easier to dispose of and safer.
12:13 To compare the two HPLCs, you can see the products here. The impurities that are downstream are more associated with the R groups. The green approach maintains the integrity of the R group and improves overall product yield.
13:13 We aim to create greener approaches by omitting the use of DMF, diethyl ether, and TFA. This was mainly based on the need to keep the R group intact, and this has set a precedent to create greener approaches.
13:17 Diego will now talk about our new manufacturing facility and symposium.
13:28 Thank you, Chris.
13:32 I will now go over the corporate presentation of our company.
13:38 The founder is Doctor Shawn Lee, a highly respected peptide chemist. He previously founded American Peptide Corporation, which later became part of Bachem.
14:05 Our team consists mainly of peptide chemists, except for Doctor Fong, our oligonucleotides director.
14:27 This is the BD team in the US, with members on the East Coast, West Coast, and internationally.
14:46 We were founded in 2001 and were the first peptide CDMO audited by the FDA in Asia.
15:07 We have been audited by various regulatory bodies, including the EMA, Japanese, Chinese, Korean, and Brazilian authorities.
15:15 Our team is divided into production, QA/QC, and regulatory areas.
15:36 We have three locations: Hangzhou, China, where we do manufacturing; San Jose, our US headquarters; and a new facility in Rocklin, CA.
16:07 The Rocklin facility will be strictly GMP and will handle everything from proof of principle to commercial batches.
16:24 Our Hangzhou campus has 24 process suites for production and 19 purification suites, with a capacity of up to 500 kilos per year.
17:07 We offer various products and services, including custom peptide synthesis, FTE services, R&D, and GMP manufacturing.
17:47 We mainly do solid phase but we also have a hybrid approach which combines solid phase and solution phase. We specialize in difficult peptides and modifications with several in the clinic. We have expertise in pegylation and FTE services. The FTE services are very popular, this involves hiring a group of chemists to work on a project for R&D purposes, this is very effective if you have lead optimisation of your peptide.
19:03 We can take you from proof of principle discovery to commercial batches, providing CMC documentation and everything needed for a GMP campaign.
19:38 We have customers worldwide, producing commercial batches for nine commercial customers and various customers in different clinical phases.
20:04 We also offer neoantigen peptides and have suites dedicated to immunotherapy.
20:19 Our largest vessel is a 1000-litre reactor, and we are growing, with plans to purchase a 30-inch prep column.
20:34 We have two project managers: an account manager and a lead chemist, who work closely with customers.
21:15 We also produce generics, which is a significant part of our business.
21:48 Our generics portfolio is broad and some have DMFs and some are going to file for DMFs. Luprolite is our flagship compound which we offer in commercial batches for prostate cancer.
21:56 We offer a symposium on peptide therapeutics, inviting customers, regulatory people, and venture capitalists.
22:39 The symposium includes three days of meetings and a tour of our manufacturing facilities.
23:13 During COVID, we offered a popular webinar on peptide therapeutics.
23:23 Thank you, Oxford Global, for inviting us and giving us the opportunity to give a talk.
