Cell Cycle Synchronous Chemotherapy
Optimizing Chemotherapy by Tumor Specific Phase Enrichment, Phase Synchronization, and Meeting Requirements for Achieving Curative Outcome.
Slide 1: Opening Slide
CELL CYCLE SYNCHRONOUS CHEMOTHERAPY
(CCSC protocols)
Presentation at Stanford Genome Technology Center
Presentation of September 2, 2004, Updated 2006, 2007
by Mark Zamoyski
e-mail: zamoyski@metricmail.com
© 2004-2007 Mark Zamoyski & NexGen Biomedical, Inc. , all rights reserved
Slide 2: Seminar Synopsis
• The Slide Show combines molecular biology, mathematics, and cytokinetics of tumor growth, kill back, and regrowth to reveal why today’s chemotherapy fails to cure most cancers.
• The seminar identifies existing drugs that can be used to prevent this failure.
• potential “fast cure” for more than a third of cancers,
including breast and prostate cancer
• “bloodless surgery” for non malignant growths,
including endometriosis and enlarged prostate
Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 3: Cancer Status / Stats
• Surgery provides 80% of today’s survival benefit.
• Leaves remaining 20% from:
• Radiation Therapy
• Chemotherapy
• Other
• Today’s Chemotherapy is not curative.
• It only extends survival by a few months of misery.
• Stage IV survival rates:
• 1% lung cancer
• 5% colon cancer
• 14% breast cancer
Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 4: Molecular Biology of Cancer
• Cancer is the accumulation of several independent genetic accidents (mutations) in growth control pathways in a single aberrant cell.
• That aberrant cell, and all of its progeny, are hard wired to relentlessly grow and divide (cycle).
• Growth Control Pathways are used for population density management
• Cells produce and release protein growth factors (~ 50 known)
• Known causes of mutations:
• Cells also possess combinations of growth factor receptors
• Cells also produce proteins that inhibit growth
• Population Density is a balance between stimulators and inhibitors of growth
• In Cancer, mutations tip the balance in favor of growth
• DNA replication errors during cell division (~ 3 per division)
• Chemical Carcinogens (DNA damage)
• Electromagnetic radiation (covalent bond breakage)
• Roughly 15% of cancer mutations trace back to viral origins
Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 5: Growth Control Pathways (1 of 3)
• Growth factor docking with a receptor triggers intracellular cascades that result in activation of the Cell Cycle Control System . Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 6: Growth Control Pathways (2 of 3)
• The Cell Cycle Control System is driven by production and combination of Cyclin/CDK proteins. Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 7: Growth Control Pathways (3 of 3)
• The Cyclin/CDK complexes activate transcription factors for cell growth and division. Return to Slide Show Index
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 8: Examples of Common Mutations
• Mutations can occur anywhere along growth control pathways. • Overexpression of growth factor receptors (slide 5)
• RAS transduction molecule malformed to be “always active” (slide 5)
• Rb gene frequently missing in lung, breast, and bladder cancers (slide 7). • About 50% of cancers have defects in the P53 gene (slide 7)
Return to Slide Show Index • ~ 50 chemotherapeutic agents available for use today • Most chemotherapeutics used today are Cell Cycle Active Cytotoxic
• The phase distribution for a typical cycling human cell is shown below Return to Slide Show Index • “Dose limiting toxicity”
• “Maximum Tolerated Dose”
• Bone marrow toxicity is dose-limiting for most chemotherapeutics • From a molecular biology perspective, for a phase specific chemotherapeutic:
Return to Slide Show Index ( per Harrison’s Principles of Internal Medicine, 14th ed. P. 528) • Cancer growth is logarithmic
• Tumor Kill Back: Each administration of chemotherapy kills a constant percentage of the tumor
Return to Slide Show Index • To be curative, you must get below the 1 surviving cell number
• Computing bone marrow recovery under Skipper:
• S-Phase Cytotoxics and Skipper:
Return to Slide Show Index • A tumor follows a Gompertzian growth curve (not linear) • Density Related Stasis 1: Pressure restricts blood flow impairing nutrient and oxygen delivery
• Density Related Stasis 2: Upregulation of density dependent inhibition pathways in cancer cells (e.g. P27 on slide 7)
• Density Related Stasis 3: Ambient Growth Factor (GF) depletion / progressively unfavorable GF/GFR ratio
• Endocrine Related Stasis
Return to Slide Show Index • Gompertzian Growth implies heterogeneity of cell cycle times in a tumor (PDT is an average) • Gompertzian Curve implies that chemotherapy itself alters cycle time by “Gompertzian Acceleration”
Return to Slide Show Index • Used Irinotecan Phase III colon cancer trials (7 day AIs, 4 administrations per cycle) • Results - Wall of numbers (Table 2, Pat. 6,486,146), cancer population cell count progression by treatment arm, shown below:
• Results - compared to actual median survival, shown graphically below:
Return to Slide Show Index • Corrected Model for conventional, asynchronous regimen and 10 bil. cell tumor mass Return to Slide Show Index • Can only expect survival increase of ~ 1/2 PDT per administration of S-Phase cytotoxic (when PDT < AI) Return to Slide Show Index • Objectives of the CCSC Protocols Return to Slide Show Index • Endocrine Dependence (ED) is a trait inherent in the cell type from which the cancer arose
Return to Slide Show Index • Step 1): S-Phase Cytotoxic + Anti Cytostatic used to de-populate the tumor and remove density related stasis points
• Step 2): Cytostatic used to limit regrowth / re-establishment of stasis and to aggregate survivors where desired
• Step 3): Anti Cytostatic used to release cells into the S-Phase
• Step 4): An S-Phase Cytotoxic is used to kill the released cells • Steps 2 - 4 are repeated to re-aggregate survivors and kill them, progressively reducing the tumor to where all cells will be cycling and available for killing Return to Slide Show Index • One Cytotoxic administration = 1 Skipper cycle
• Population asynchronicity is removed by phase aggregation
• Gompertzian acceleration does not result in asynchronicity, cells just get to the road block faster (i.e. the synchronization point)
Return to Slide Show Index • Maximum tolerated dose is replaced by minimum efficacious dose
• Uses cancer accelerants for further dose reduction
• Today’s Maximum tolerated dose is a bad tradeoff
Return to Slide Show Index • 184,200 new cases of breast cancer
• 180,400 new cases of prostate cancer
• 36,100 new cases of ovarian cancer
• Same principles could provide “bloodless surgery” for other growths
Return to Slide Show Index • Molecular biology indicates receptor blockers (antibodies) would be cytostatic (slide 5) • Molecular biology predicts antagonistic function of trastuzumab and cell cycle active cytotoxics, when used as it was in Phase III human clinical trials
Return to Slide Show Index Patent applications are currently pending in this area and a signed NDA is required prior to disclosure. The website slides will be updated as appropriate.
Return to Slide Show Index Patent applications are currently pending in this area and a signed NDA is required prior to disclosure. The website slides will be updated as appropriate.
Return to Slide Show Index Patent applications are currently pending in this area and a signed NDA is required prior to disclosure. The website slides will be updated as appropriate.
Return to Slide Show Index Patent applications are currently pending in this area and a signed NDA is required prior to disclosure. The website slides will be updated as appropriate.
Return to Slide Show Index • Curative result cannot be expected from today's 7 or 21 day regimens
• ~ 10 % of cancers do not over express telomerase • Higher (14%) breast cancer survival likely due to ED related S-Phase crawl Return to Slide Show Index • Curative result cannot be expected from today's 7 or 21 day regimens • “Fast Cure” for many cancers potentially already available • No matter how magnificent of an anti cancer molecule you develop, it will fail to work magnificently if not administered properly Return to Slide Show Index
DISCLAIMER AND IMPORTANT NOTICE: The Compositions and Methods presented on this website are all in preclinical trial stages. They are based only on our understanding of the proposed underlying mechanisms of action and on any available coincidental corroborative empirical evidence, any of which may in fact turn out not to be correct, or may be prevented from functioning as envisioned because of other factors or mechanisms of action not contemplated or considered, or may even cause harm because of factors or mechanisms of action not anticipated. The process of obtaining FDA approvals has not been started in any of the areas disclosed on this website. The disclosures here are purely for scientific information exchange purposes, representing one scientific point of view, and are not intended to suggest, or be used for, any proposed medical treatments. © 2002 - 2007 Mark Zamoyski & NexGen Biomedical, Inc.
• Cell preferentially takes up more ambient growth factors
• Intracellularly mimics elevated level of growth factors
• Example: HER2 (EGF receptor) overexpressed in ~ 25% of breast cancers, 30% of NSC lung cancers, and in ovarian cancer.
• Example: EGFR overexpressed in ~ 50% of glioblastomas (brain cancer).
• Transmits false grow signal to nucleus
• Found in ~ 25% of cancers
• P53 prevents cell division or kills cells with DNA damage
• Its absence allows genetic mutations to accumulate
• Many chemotherapeutics rely on P53 for their cytotoxic affect
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 9: Chemotherapy Overview
• Typically 7 or 21 day administration intervals (AIs)
• Typically 4 - 12 administrations per regimen• most are S-Phase Cytotoxic
• The S-Phase is the part of the cell cycle when DNA is being Synthesized
• Cells in or passing through the susceptible phase are killed
• Chemo kills both cancer and normal actively cycling cells
• "The Big 4" normal actively cycling cells: bone marrow, gastrointestinal, hair, and skin
• Frequent turnover cells include ovaries, testes, thymus, lymph nodes and spleen
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 10: Principles of Chemotherapy - Definitions • Toxicity to normal tissue that limits further dose escalation
• The dose just below the “Dose limiting toxicity”
• max. tolerated dose equals max. systemic toxicity
• max. tolerated dose does not equal an appreciable increase in tumor kill rate (TKR)
• max. tolerated dose does not equal curative result
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 11: Principles of Chemotherapy - Skipper Log Cell Kill Model (1 of 2)
• Cells / Population Division: 1, 2, 4, 8, 16, 32, 64, 128, 256, 512, ...
• To be curative, you must get below the single surviving cell number
• No growth can occur between administrations
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 12: Principles of Chemotherapy - Skipper Log Cell Kill Model (2 of 2)
• For a 10 bil. cell tumor, need 6 cycles for a 99% TKR/cycle chemo.
• 10 bil. X .01 survivors X .01 X .01 X .01 X .01 X .01 < 1
• For a 10 bil. cell tumor, need 8 cycles for a 95% TKR/cycle chemo.
• i.e. need to multiply 10 bil. by .05 eight times to get < 1
• e.g. 99% bone marrow cell kill = 1% survivors
• from 1% to 2% to 4% to 8% to 16% to 32% to 64% to 100% =~ 7 population divisions
• at a 24 hr. cycle time, = ~ 7 days to normal population density
• A 99% S-Phase Kill Rate (PKR) =~ 32% TKR (tumor kill rate)
• Need 4 administrations of the chemotherapeutic, synchronous to the progression of tumor cells through the S-Phase, for 1 Skipper cycle
• No cells may slip past the S-Phase before the next administration of chemotherapy
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 13: Gompertzian Tumor Growth
example for a 4 day “hard wired” cycle time
• Population division time(PDT) is 5X slower above the 1cc mass
• Low S-Phase fractions are often observed (e.g. 10 -15% vs. 32%) 
Tumor Density Related Stasis
• directly counteracts all mutation profiles, across all phases of the cell cycle
• also causes angiogenesis inhibition by P27 upregulation in endothelial (blood vessel) cells
• manifests as slowed tumor growth
• Tips balance back in favor of stasis (i.e. not growing)
• Counteracts GF, GF Receptor, transduction and transcription mutations in the G1 phase
• Would manifest as slowed tumor growth with low S-Phase fraction
• Growing tumor counteracts GF Receptor overexpression mutations
• GFR overexpression relies of preferential uptake of ambient growth factors
• Growing tumor and fixed ambient growth factor pool leaves unbound / unactivated receptors
• Counteracts GF Receptor overexpression (in the G1 phase)
• Would manifest as slowed tumor growth with low S-Phase fraction
• Endocrine dependent cancers (e.g. estrogen, progesterone, testosterone) require endocrine binding for entry into and progression through the S-Phase
• estrogen and progesterone levels vary through the menstrual cycle and nadirs result in stasis
• chemotherapy causes ablation of ovaries and testes with resulting downregulation of estrogen and testosterone
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 14: Gompertzian Growth, Asynchronicity, and Why Chemotherapy Fails
• the stasis points (slide 13) would be most pronounced in the dense core, least pronounced at the periphery
• cells in the dense core would cycle slower than at the periphery
• also, cells next to a blood vessel would cycle faster than ones 6 lengths away
• heterogeneity of cycle times implies inherent asynchronicity to 7 or 21 day administration intervals (AIs)
• without synchronicity, curative result cannot be expected under the Skipper model• Chemotherapeutic tumor de-population removes the density related stasis points (slide 13)
• cells accelerate their cycle time until they recrowd
• leaves them asynchronous to the regimen
• without synchronicity, curative result cannot be expected under the Skipper model
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 15: Clinical Corroboration of Chemotherapeutic Asynchronicity
• Used standard S-Phase fraction of 32% and 100% S-Phase kill rate (= 32% tumor kill rate per administration)
• asynchronicity of the S-Phase in the cancer cells to successive administrations of S-Phase cytotoxic would mean only a modest increase in life expectancy as the cancer kept regrowing
• the increase in life expectancy could be approximated using Skipper log cell kill math: regrowth # = (starting # of cells) X 2 (T / PDT in days)
• where T is days between administration of chemotherapy
• where a clinically detectable 1cc mass (1 bil. cells) was used as the start point
• where an average lethal burden mass of 1 liter (1 trillion cells) was used as the end point
• and where the PDT (Population Division Time) was estimated as 19 days based on the best supportive care (BSC) life expectancy (ie. BSC / 10 population divisions)
• Clearly not curative
© 2002 - 2007 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 16: Zamoyski Corrected Model of Tumor Kill Back and Regrowth
• based on Irinotecan clinical trial observations (i.e. formula of slide 15)
• only a delay in progression to lethal burden can be expected
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 17: Simplified Explanation
• S-Phase chemo kills ~ 1/3 of the tumor per each administration
• The surviving 2/3 needs to grow ~ 50% to restore the original tumor size.
• This only delays the tumor’s progression to lethal burden by 1/2 population division per administration
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 18: Overview of the CCSC Protocols
• Make Chemotherapy Curative
• Conformed Cancer Concepts (Pat. 6,486,146 and others pending)
• Insure Synchronicity of cytotoxic administrations to the susceptible phase
• Minimize Systemic Toxicity
• Conformed Cancer Concepts are used to insure synchronicity
• Minimum Efficacious Dose Concepts
• Cancer Accelerant Concepts• The process synchronizes the tumor’s susceptible phase to administrations of a phase specific cytotoxic
• The regimens typically start with a de-populating administration(s) to remove density dependent stasis (slide 13)
• Cytostatics are used to prevent re-establishment of density dependent stasis and to phase aggregate tumor cells where desired
• Anti-Cytostatics are used to release the aggregated cells
• Cytotoxics are used to kill the phase aggregated, released cells
• The process is repeated to re-aggregate and kill survivors
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 19: CCSC Protocols for Endocrine Dependent Cancers (EDC)
( Pat. # 6,486,146 and other applications are pending)• It is a trait retained by the endocrine dependent cancer (EDC)
• Endocrine receptors are formed during the G phase
• Endocrine (e.g. estrogen, testosterone) binding to these receptors is required for entry into and progression through the S-Phase
• Endocrines can be used as Anti-Cytostatics
• Absence of ED receptors in big 4 = tumor specificity for chemotherapy purposes
• Provides a method for selectively synchronizing the cancer to the chemotherapy
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 20: CCSC Protocols for Breast Cancers
( Pat. # 6,486,146 and other applications are pending)
• Numerous S-Phase cytotoxics available (Harrison’s, 15th ed. P. 539 - 540)
• cytostatics include estrogen downregulators such as aromatase inhibitors
• Anti Cytostatics include exogenous estrogen, estradiol, etc...
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 21: CCSC Protocols and the Skipper Kill Model
• PKR = TKR in a conformed tumor
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 22: Taking the Misery Out of Chemotherapy• Astronomical increase in normal population kill back for an insignificant increase in tumor kill back
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 23: Applicability of CCSC Endocrine Conformed Regimens
( Pat. # 6,486,146 and other applications are pending)• ~ 2/3 are estrogen dependent
• ~ 1/3 are progesterone dependent• most are testosterone dependent
• administer as PSA levels start rising to obviate surgery / prevent eventual malignancy• many progesterone dependent
• endometriosis (estrogen dependent
• enlarged prostate (testosterone dependent)
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 24: Incorrect Cytostatic Regimens - Trastuzumab
• ~ 25% of breast cancers over express HER-2 receptors
• trastuzumab is a HER-2 antibody (e.g. Herceptin, a registered trademark of Genentech)
• it should prevent docking of ambient growth factors with overexpressed receptors
• it should result in G-Phase stasis
• Trastuzumab induced G-Phase stasis clinically corroborated by Bunn et. al.
• trastuzumab was administered for 4 months and cell cycle active cytotoxics were administered at 21 day intervals during that time period
• The cytostatic would prevent cancer cells from cycling
• A Cell Cycle Active Cytotoxic requires cells to be cycling in order to kill them
• Accordingly, the first administration of the M or S-Phase Cytotoxic would be expected to work as cells were still progressing through the M or S Phase
• However, the next 5 administrations of the M or S-Phase cytotoxic would be expected to only cause systemic toxicity, with no commensurate therapeutic benefit, as surviving tumor cells were arrested in the G-Phase
• Per Skipper log cell kill math, one would only expect the benefit of ~ 4 mo. of cytostatic + 1 asynchronous cytotoxic kill back
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 25: Clinical Corroboration - Skipper Math and the Phase III Data
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 26: Corrected Conforming Regimens using Trastuzumab
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 27: Regimens for Non Conformable Cancers (1 of 2)
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 28: Regimens for Non Conformable Cancers (2 of 2)
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 29: Closing Comments / Tie Up of Loose Ends• Then why the Sage IV survival rates of 1% for lung cancer or 5% for colon cancer?
• Cancers not over expressing telomerase would be subject to cell senescence
• the average cell has 50 - 60 cell divisions before telomere reduction results in cell senescence
• It takes ~ 40 cell divisions to reach lethal burden
• Depending on how many cell divisions were used up by the cell before it went malignant, plus how many population divisions you force the cells to use up because of chemotherapy, you may hit senescence before lethal burden
• Would expect between 0 - 10% survival from senescence and the observed 1% and 5% is in this range
• Other possibilities: slow mutation profiles, whose fastest rate may coincidentally match the AI
• Chemotherapeutic ablation of ovaries in ~ 71% of women
• Ablation = estrogen downregulation = S-Phase crawl
• Haphazard phase aggregation / Gompertzian acceleration offset
• Similar, in part, to what we do much more precisely
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 30: Conclusions / Implications
• An expectation corroborated by abysmal Stage IV survival rates
• at a minimum, high tumor response rate, low toxicity regimens
© 2002 - 2006 Mark Zamoyski & NexGen Biomedical, Inc., all rights reserved
Slide 31: Speaker Bio
Mark Zamoyski is the principal scientist and founder of NexGen Biomedical. Mark was awarded a patent for cell cycle synchronous chemotherapy for endocrine dependent cancers and has applications issued and pending for tumor specific, cell cycle synchronous protocols for several other cancer characteristics and mutations. Mark also has patents issued and pending for RNAi / PSR compounds for inhalable and topical anti-viral, anti-inflammatory, and antiproliferative treatments for various dermal and pulmonary indications. Mark's more recent work involves identifying a novel etiology / pathogenesis underlying many types of migraines and seizures and accordingly novel etiology based treatment methods. Prior to founding NexGen Biomedical, Mark spent more than a decade with Biomation Corporation and its parent company, where he worked his way up to executive positions including CFO and VP of Sales and Marketing and was involved in the ultimate sale of the company. Mark received both his bachelors and masters degrees from Cornell University in 1977 and 1978, respectively.
Acknowledgments: We wish to thank Dr. Maria Zagorski and Justin John Zamoyski for their help in the preparation and review of the above presentation material.