NorthWest Biotherapeutics Inc (NWBO)

[Rkmatters]

UCLA did a DC GBM grant study.

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=116779774

From there UCLA generated patents, which they licensed out to NWBO. NWBO essentially combine with patents of their own, but they used the manufacturing process that UCLA was contracted to find.

Agreement between NW Bio and UCLA went into place in April 2001,

Part of the 2001 - 2002 Regents AGREEMENT is to develop IND #10206:

3. CONTENT OF WORK:

A. Develop and validate a protocol for preparing GMP-quality suspensions of viable glioblastoma cancer cells from patients with glioblastoma multiforme and technology developed technology to Sponsor. University investigators will collect cancer specimens from the operating room and evaluate techniques for preparing purified tumor cell suspensions from these clinical samples. The goal will be to develop a GMP-quality process by which tumors recovered at the time of surgery can be placed into a transportation media and delivered to the Sponsor's cell processing facility in the form of a viable cell suspension. In addition, techniques for purifying, characterizing and culturing the tumor cell suspension after its arrival at the Sponsor's facility will be investigated. Techniques for "stripping" peptides from the surface of tumor cells and concentration of these peptides will also be developed. Results from these investigations will be formed into a detailed written protocol that will be delivered to the Sponsor. This protocol will employ GMP-quality reagents, as feasible given their current availability. The average viability, cell yield and purity of the cancer cell suspension as well as quantities of peptides "stripped" from the tumor cell surface will be reported to the Sponsor.

And this is the method they came up with:

"Manufacturing process:

The tumor is excised by a neurosurgeon and a portion thereof removed for pathological examination. All remaining tumor tissue is diced and digested to a single cell suspension using an enzyme mix. Next, the tumor cell suspension is filtered through a strainer and then centrifuged for 10 minutes to pellet the tumor cells. The aspirate is removed and the cell pellet is re­suspended in RPMI­1640 without phenol red, and transferred to cryovials previously labeled with patient ID information. The cell suspension in cryovials is then subjected to freeze­ thaw cycles to effectively lyse the tumor cells. The tumor lysate is pooled and centrifuged.The supernatant is then filtered through a 0.2 µM filter. Sealed vials containing the tumor lysate are frozen and stored at -80C in controlled inventory until used for loading DC generated from the patient.

Dendritic Cell Preparation Peripheral blood mononuclear cells are isolated by leukapheresis from the patient before treatment begins. MNC are purified by density gradient centrifugation, and either used fresh or cryopreserved until further processing. After thawing of the MNC, if cryopreserved, adherent cells are cultured ex vivo for 6 days with IL­4 and GM­CSF to generate DC. The DC are collected and combined with the tumor lysate antigen for 16 hours to prepare DCVax­ L. After the incubation, the final product is harvested and cryopreserved for shipment to the clinical site for patients who are randomized to the treatment cohort. " - IND 10206 (developed in 2001-2002)

IT speaks nothing about UCLA or NW Bio's patents only of the agreement to which around GMP suspension. Nothing around pulsing lysate which we know is done. And now that I pointed out his assumption on when the IND was created, which he said was late 1990s, I've asked him how he could easily discount and ignore my arguments that patents from NW Bio and UCLA that would be incorporated into this Phase II (now III) as they specifically address why additional maturation agents would not be NEEDED in the manufacturing if they are included within the combining DC and antigens.

Today he stated the techie is old, methods are old. But, After Phase I, UCLA was looking into other methods for DC pulsing (combining with antigens):

"The number of passages required for obtaining sufficient peptide for each patient varied from 3 to 12. Thus, vaccine administration was sometimes delayed by the time required to obtain sufficient peptide antigen (i.e., >4 weeks). For this reason, we are concurrently investigating other methods of dendritic cell pulsing, the clinical results of which will be subsequently compared with those reported here. — Linda Liau after Phase I"

Patents that specifically address this "The DC are collected and combined with the tumor lysate antigen for 16 hours to prepare DCVax­ L. " and "After the incubation, the final product is harvested" include:

Methods for detection and treatment of neural cancers
Patent number: 6558668

Abstract: The invention provides a method for inhibiting proliferation of neural cells. The neural cells can be tumor cells, glial cells, neuronal cells, and cells of the central or peripheral nervous systems. The method comprises contacting a neural cell with a molecule that disrupts the biological activity of a granulin molecule. In one embodiment, the molecule is an antibody directed against a granulin peptide. In other embodiments, the molecule is an antisense nucleotide directed against a granulin nucleic acid molecule, or a vaccine comprising a granulin peptide or a polynucleotide encoding a granulin peptide. The invention additionally provides methods for detecting and treating cancer in a neural tissue using granulin-related molecules. Also provided is a method for identifying differentially expressed gene products that are translated from mRNA species, using antibody-based screening of a cDNA expression library.
Type: Grant
Filed: February 28, 2001
Issued: May 6, 2003
Assignee: The Regents of the University of California
Inventor: Linda M. Liau

This application claims benefit of U.S. provisional application No. 60/185,321, filed Feb. 28, 2000, the entire contents of which are incorporated herein by reference. Throughout this application various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to describe more fully the state of the art to which this invention pertains. Some of these references are indicated by numbers in parentheses. Citations corresponding to these reference numbers can be found at the end of the specification

A pharmaceutical composition or vaccine can contain DNA encoding one or more of the polypeptides as described above, such that the polypeptide is generated in situ. As noted above, the DNA may be present within any of a variety of delivery systems known to those of ordinary skill in the art, including nucleic acid expression systems, bacteria and viral expression systems. Numerous gene delivery techniques are well known in the art, such as those described by Rolland, Crit. Rev. Therap. Drug Carrier Systems 15:143-198, 1998, and references cited therein. Appropriate nucleic acid expression systems contain the necessary DNA sequences for expression in the patient (such as a suitable promoter and terminating signal). Bacterial delivery systems involve the administration of a bacterium (such as Bacillus-Calmette-Guerrin) that expresses an immunogenic portion of the polypeptide on its cell surface or secretes such an epitope.

When DCs are pulsed with a soluble antigen, including human tumor antigen or tissue specific antigens with an adjuvant such as BCG, enhancement of MHC-class I presentation occurs. Therefore, the presence of an adjuvant such as BCG typically increases DC soluble tumor antigen processing in the MHC-class I compartment and correspondingly, activates a higher percentage of CD8+ T cells when compared to individuals administered the antigen alone. "- UCLA patent

NW Bio patent (2001) of combining DC with the tumor lysate antigen for 16 hours to prepare DCVax­ L. - IND 10206 (developed in 2001-2002):

https://docs.google.com/viewer?url=patentimages.storage.googleapis.com/pdfs/US20050059151.pdf


Sep 6, 2001

Compositions and methods for priming monocytic dendritic cells and t cells for th-1response
US 20050059151 A1

The present invention provides compositions and methods for inducing maturation of immature dendritic cells (DC) and for priming those cells for inducing a type 1 immune response. The present invention also provides dendritic cell populations useful for activating and for preparing T cells polarized towards production of type 1 cytokines and/or a type 1 response. Similarly, activated, polarized T cell populations, and methods of making the same are provided."
+

"The tumor cell can be prostatic, lung, ovarian, colon, BRAIN, melanoma, or any other type of tumor cell. " -- Marnix Bosch patent

And so yes, I concluded that BCG and INFy are most definitely used in the formulation. :)