Technology behind Lipoplatin


Regulon is an oncology-focused drug delivery company founded in Silicon Valley, California. Its mission is to alleviate symptoms of human disease, especially cancer, by applying liposomal nano-encapsulation technologies to existing cancer chemotherapy drugs. Its first product is a liposomally encapsulated cisplatin nanoparticle developed by Dr. Teni Boulikas in 1999 in California. This product is called Lipoplatin as an orphan drug against pancreatic cancer and Nanoplatin for its application to Non-Small Cell Lung Cancer (NSCLC). Cisplatin is the most widely- used chemotherapy drug, the gold standard for epithelial malignancies with applications to over 50% of human cancers.
The virtues of Lipoplatin nanoparticles include:
(i) passive targeting of tumors and metastases. Lipoplatin nanoparticles, loaded with cisplatin are uptaken by tumors and metastases by leaking through the compromised endothelium of tumor vasculature sprouted during neoangiogenesis, a process known as extravasation, and by the avidity of tumors for nutrients with Lipoplatin disguised as a nutrient with its lipid shell. The passive targeting results in concentrations of Lipoplatin in tumors that are 10-200 times higher compared to the concentration of Lipoplatin in the adjacent normal tissue from human studies. It takes approximately 12 to 24h to reach this optimal concentration into tumors after intravenous infusion of the drug to patients. The endothelium of normal tissue does not allow Lipoplatin nanoparticles to extravasate and thus the drug concentrates in tumors rather than normal tissue. Thus, Lipoplatin treatment causes no side effects. In fact at doses 5 times higher than the dose of cisplatin in a monotherapy study against NSCLC only Grade 1 toxicities were reported compared to Grade 3 and 4 by the vast majority of other chemotherapy drugs. The response rate in patients with NSCLC was 38% with Lipoplatin compared to 11% with cisplatin from historic controls.
(ii) antiangiogenesis potential by attacking and destroying endothelial cells of tumor vasculature as well as epithelial cells in contrast to cisplatin that destroys only epithelial cells
(iii) ability to fuse with the cell membrane and thus to deliver its toxic payload, cisplatin, in the cytoplasm of tumor cells. The ultimate result is the delivery of the most powerful cisplatin to the cytoplasm and nucleoplasm of tumor cells and to the cells of their vascular endothelium. This bypasses resistance at the cell membrane barrier and results in a higher cytoplasmic concentration of the drug causing massive destruction to tumor cells by intra-strand DNA crosslinks, signaling modulation, mitochondrial apoptosis and by increasing the production of destructive free radicals in cancer cells.


(iv) ability to absorb energy from external sources, such as radiation of patients treated with Lipoplatin the previous day, a process that enhances the reactivity of the heavy-metal containing Lipoplatin nanoparticle. It is like the heating of a metal but not of a wooden sphere by applying the same thermic energy on them.
(v) Lipoplatin is endowed with antimetastasis properties thus the metastases are minimized during treatment of the patients.
Published Phase III studies (see Phase III results) show a lower toxicity with improved therapeutic index in adenocarcinomas of the lungs compared to cisplatin; this feature was not shown by other platinum blockbuster drugs (carboplatin, oxaliplatin). In a randomized Phase III in NSCLC, a statistically significant reduction of neutropenia, nephrotoxicity and asthenia of cisplatin was demonstrated by its replacement with Lipoplatin. Also in a randomized Phase III in nonsquamous-NSCLC the partial response for the Lipoplatin arm was 59% compared to 42% for the cisplatin arm and the difference was statistically significant (p=0.036).

Effective cancer treatment with Liposomal Cisplatin monotherapy and low-dose radiation

The most recent highlights of Lipoplatin include its application to many different cancer indications as monotherapy in combination with low-dose radiation. Patient case reports with osteosarcomas, lung, breast, kidney cancer, glioblastomas, leukemias and other cancers (manuscripts in preparation) demonstrate a highly effective treatment void of side effects that brings a true revolution in the cancer field. The success of response in patients irrespective of cancer indication, at stage IV of their disease, is almost

100%. The protocol involves treatment with Lipoplatin monotherapy using 200 mg/m 2 on Day 1 followed by 100-200 mg/m 2 on Day 2 (depending on age, performance status) followed by radiation with 2-3 Gray on Day 2. This

treatment is repeated every week for 2-3 months. The patient is subject to

PET/CT before and after treatment to follow progress.

Mission of Regulon, Inc.

Regulon is a private biopharmaceutical company founded in Silicon Valley, California. Our mission is to alleviate symptoms of human disease, especially cancer, by applying liposomal nano-encapsulation technologies to existing cancer chemotherapy drugs. Regulon is possessing a world-wide patented nano-technology that has been applied to cancer chemotherapy achieving important goals and solving major and outstanding problems in clinical oncology: Effective passive tumor targeting, transport of the toxic payload across the cell membrane barrier, lowering of the side effects of the classical chemotherapy, synergy with radiation for tumor cell killing. Finally, Lipoplatin nanoparticles are endowed with antiangiogenesis and antimetastasis potentials, desperately sought by oncologists among drugs, all combined together in the same nanoparticle along with the classical chemotherapy virtues of cisplatin.

Technology & Mechanisms in detail:

1. Effective passive tumor targeting

Passive delivery to tumors is achieved secretly from immune cells and normal tissues by encapsulation of the cytotoxic drugs into a natural lipid capsule protected with a PEG polymer; the 110-nm liposome nanoparticles exploit the compromised

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endothelium of tumor vasculature for their preferential extravasation to tumors and metastases. Lipids, composing the nanoparticle shell, are natural products, one of the four classes of biomacromolecules, compatible with the lipids of the cell membrane, unlike synthetic polymers used in other nanotechnology capsules with dubious cumulative toxicities. Regulon’s nanoparticles carrying therapeutic drugs have long circulating properties in body fluids (Figure 1); this longevity in circulation is required for passively identifying the tumors and metastases in the body for their preferential targeting and extravasation in tumors. This preferential targeting of cancer tissue takes advantage of the imperfections in the vasculature sprouted by tumors to accelerate their growth during neoangiogenesis; the arteries, veins and micro-vessels in normal tissue have endothelial walls more compact compared to the “leaky” vasculature of tumors; as a result, tumors uptake 10- to 200-times more Lipoplatin nanoparticles than normal tissue (Figure 2); this was demonstrated in human studies where patients were infused with Lipoplatin and the platinum levels were measured in surgical specimens from primary or metastatic tumors and the adjacent normal tissue. Regulon’s anticancer treatment minimizes the side effects of classic chemotherapy. In simple terms all primary tumors and metastases are being targeted regardless of the tumor type or size following intravenous administration of our drug. Their targeting depends primarily on the degree of tumor vascularization. Tumors of the stomach and breast for example have the highest degree of vascularization and are expected to accumulate more platinum drug after intravenous administration.

Figure 1. The scheme shows the PEGylated liposome that is the carrier of the toxic drug cisplatin with its long-circulating properties in body fluids after intravenous administration.

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Figure 2. The process of Lipoplatin extravasation. The scheme shows a blood vessel in tumor tissue. Lipoplatin nanoparticles of 100nm in diameter are depicted as spheres with the yellow toxic payload of cisplatin inside them. In normal tissue, blood vessels are impenetrable by small nanoparticles. On the contrary, tumor blood vessels have imperfections (tiny holes) in their walls (called endothelium); tumor blood vessels are established during the process of neo-angiogenesis (meaning sprouting of new blood vessels by a tumor cell mass during its growth phase). Lipoplatin nanoparticles take advantage of these tiny holes to pass through and extravasate inside the tumor reaching a concentration that can be 10- to 200-fold higher compared to the adjacent normal tissue.

2. Crossing of the cell membrane barrier by Lipoplatin nanoparticles leading to delivery of their toxic payload inside the cytoplasm of the tumor cell where it is needed for anticancer efficacy (Figure 3). This is a major advantage in the implementation of the treatment in the clinic to enhance efficacy and eliminate toxicity. Crossing of the cell membrane barrier by Lipoplatin was also demonstrated in cell cultures (Figure 4).

3. Lowering of the side effects of the classical chemotherapy. Because of the lipid shell, Lipoplatin does not harm the cells of the kidney and other normal tissues to cause nephrotoxicity and other side effects. On the contrary, because of its extravasation (Figure 2) and its penetration inside the cell by fusion with the cell membrane (Figure 3) it enhances efficacy while lowering penetration into normal tissue thus lowering side effects of classical cisplatin chemotherapy (toxicity to kidneys, bone marrow, peripheral nerves, gastrointestinal tract).

Lipoplatin is rapidly phagocytosed by tumor cells bypassing the membrane barrier which is largely responsible for drug resistance which commonly arises with 1st line therapy
Unmodified “naked” cisplatin uses copper transporter 1 (Ctr1) receptor mediated transportation to enter tumor cells
Reduced platinum uptake is a key factor in Cisplatin resistance and cisplatin resistant cancers show a lower-level of Ctr-1 expression
DPPG mediated fusion and phagocytosis circumvents the need for Ctr1 transportation

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Thus, Lipoplatin has the potential to treat patients that have failed previous platinum-based treatment and have developed tumors resistant to platinum drugs at the cell membrane level. An additional level of platinum drug resistance is at the level of DNA repair where resistant cells can repair DNA lesions faster.

Figure 3. Delivery of cisplatin “payload” directly to tumor cells facilitated by DPPG fusion circumventing the need for Ctr1-receptor mediated transportation required by naked cisplatin. After concentrating in tumors and metastases DPPG promotes the fusion of Lipoplatin with the cell membrane. Once they reach the tumor target Lipoplatin nanoparticles have the advantage, unique to Regulon’s technology, to fuse with the cell membrane of the tumor cell and empty their toxic payload inside the cytoplasm. Liposomes developed by others (e..g. Doxil of SPI-77 of Alza/J&J) are unable to do the fusion process; thus the toxic drug is emptied outside the tumor cell and is less effective.

Figure 4. Demonstration of the fusion or uptake of Lipoplatin nanoparticles using cancer cell cultures. The green donut-like structures are single cancer cells; their periphery where the cell mebrane is located fluoresces because it has uptaken fluorescent Lipoplatin nanoparticles or Regulon’s fusogenic liposomes as a control. Lipoplatin or DPPG-liposomes with fluorescent lipids enter rapidly MCF-7 human breast cancer cells thus providing proof of concept of membrane fusion or endocytosis to deliver the toxic cisplatin inside the tumor cell.

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4. Synergy with radiation for tumor cell killing . Lipoplatin is the only nanoparticle drug available that contains a heavy metal inside a liposome. Platinum can uptake high energy from external sources such as laser or gamma rays that can burst the nanoparticle to release the toxic drug or to heat up the surrounding cytoplasm. These exciting properties are under current investigation to explore the full potential of this exciting nanoparticle.

5. Antiangiogenesis properties . Lipoplatin nanoparticles are endowed with

antiangiogenesis properties.
The antiangiogenesis property of Lipoplatin has been suggested from the encapsulation of the beta-galactosidase gene into a liposome of the same composition as the Lipoplatin liposome; after systemic delivery to SCID mice with human tumors (Figure 5) the foreign “blue” gene stained preferentially the vasculature that the tumors under the skin of the animals developed to supply the tumor with nutrients. This shows that Regulon’s liposomes can target preferentially the vascular endothelial cells; in case of Lipoplatin, targeting of these cells with toxic cisplatin instead of the “blue” gene would cause their destruction. Thus, Lipoplatin limits tumor vascularization by attacking their endothelial cells in addition to the known property of cisplatin to attack the epithelial cell of the tumor.

6. Antimetastasis potential of Lipoplatin

The antimetastasis potential of Lipoplatin was shown in a study from the “Reference Oncology Center, Italian National Cancer Institute” in Aviano. Lipoplatin inhibited both migration and invasion of cervical cancer cells supporting its antimetastasis potential. This is a very important feature of Lipoplatin because migration and invasion are essential steps used by cancers to mediate their metastases.
In this paper that appeared in September 2013 in “Gynecologic Oncology” (, the investigators, led by Dr. Donatella Aldinucci have examined the effectiveness of Lipoplatin in cisplatin- resistant cervical cancer cells. In the Aldinucci study, Lipoplatin was effective in both cervical cancer and cisplatin-resistant cervical cancer cells thus opening the possibility to apply Lipoplatin successfully against cervical cancer both as first and second-line. Furthermore, the same study has elucidated novel mechanisms on how Lipoplatin kills cervical cancer cells:
1. Lipoplatin treatment induced apoptosis in these cells, as evaluated by Annexin-V staining and DNA fragmentation, caspases 9 and 3 activation, Bcl-2, Bcl-xL down- regulation, and Bax up-regulation.
2. Lipoplatin inhibited the activity of Thioredoxin reductase (TrxR) in cervical cancer
cells. TrxR is a selenoenzyme which is over-expressed in many tumor cells and contributes to drug resistance; this finding revealed one of the several mechanisms Lipoplatin can kill cells resistant to platinum.
3. Lipoplatin induced an increase in Reactive Oxygen Species (ROS) even in the presence of the ROS scavenger N-Acetylcysteine (NAC). This is an additional mechanism for mediating tumor cell killing by Lipoplatin.
4. Lipoplatin reduced the expression of EGFR (epidermal growth factor receptor) and its phosphorylated form. EGFR is overexpressed in many tumor cells and thus the reduction of EGFR molecules on the cell membrane by Lipoplatin would have a strong anticancer effect.
5. Lipoplatin inhibited both migration and invasion of cervical cancer cells. This is a very important feature of Lipoplatin because migration and invasion are essential

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steps used by cancers to mediate their metastases; over 90% of cancer patients succumb because of complications from metastases rather than the primary tumor.

Figure 5. Encapsulation of the beta-galactosidase gene into a liposome of the same composition as the Lipoplatin and systemic delivery to SCID mice with human tumors stained preferentially the vasculature that was developed by the tumors under the skin of the animals to supply the tumor with nutrients. From Boulikas T Molecular mechanisms of

cisplatin and its liposomally encapsulated form, Lipoplatin™. Lipoplatin™ as a chemotherapy and antiangiogenesis drug. Cancer Therapy Vol 5, 349-376, 2007.

Regulon’s liposome encapsulation technology can be applied to most of the
1,000 FDA approved drugs, thus increasing the Company’s fundamental value.

Lead Product

Regulon’s lead product Lipoplatin™, is a liposomally encapsulated cisplatin. It has completed successfully Phase III clinical evaluation. Only one in 1,000 drugs tested by Pharma and Biotech Companies gets past Phase I, II and III successfully. Furthermore, although most new drugs start as second-line therapies (after failure of the recommended first-line treatment by FDA and the European EMA) Regulon’s Lipoplatin is entering the $60 Bn cancer market as first-line treatment for the worlds’s largest cancer indication, lung cancer representing over 15% of all cancer cases.
Furthermore, the same drug has been tested successfully against pancreatic, lung, gastric, breast and other cancer indications. Thus, the drug is expected to become a major player in the oncology market.
So far we have sold distribution rights to local Pharmaceutical Companies in small EU, Asian and Latin American markets while negotiating with global pharmaceutical Companies for the licensing & distribution of Lipoplatin in global markets. This is a proof of concept that Lipoplatin has successfully passed the evaluation of the pharmaceutical world.

Clinical developments

Clinical trials in Phase I, Phase II and Phase III as well many preclinical studies took place in the last decade with important publications. During this period Regulon chased its dream to develop Lipoplatin™ as first-line treatment in a big cancer indication (lung cancer, number 1 cancer in Eastern Europe, Turkey, China, most

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Asian countries) in addition to the orphan drug status in pancreatic cancer. However, it has also tested successfully Lipoplatin against breast cancer, urinary bladder cancer, and in combinations of the drug with hyperthermia, photodynamic therapy and others.
However, the biggest of the developments is cracking now after the application of Lipoplatin as monotherapy with low-dose radiation. Lipoplatin will get approval as first-line in breast and prostate cancers, the other leading cancer indications to cause a seismic cataclysm of events in the pharmaceutical industry and oncology globally. The same regimen can be applied to the vast majority of human cancers bringing, as Akesios has predicted, a true revolution in cancer treatment and making Lipoplatin the top selling drug.
Regulon is in discussions with top CROs (Clinical Research Organizations) for conducting various clinical studies mainly in EU and USA for EMA and FDA approval. However, our strategy is to include in the same studies patients from China, Japan, Canada and potential other countries in order to achieve Marketing Authorization (MA) in all these countries and globally very soon or simultaneously with EMA-FDA approval. Regulon feels that the statistical difference between the arm using Lipoplatin-Radiation compared to the Arm using the recommended treatment will be to our favor and that the number of patients required will be ~400 per study rather than 1,000 making the studies cheaper. Lipoplatin could potentially get FDA and EMA approval from a randomized Phase II like Glivek did but in a big indication.

Phase III results

Superiority of Lipoplatin to cisplatin in response rate as first-line treatment against non-squamous non-small cell lung cancer (ns-NSCLC) from a randomized Phase III study

The introduction of cisplatin in 70s has revolutionized cancer chemotherapy, especially of epithelial malignancies, most notably, testicular cancer. Carcinomas of the head and neck, bladder, ovaries, testis, esophagus, and lung are the most common malignancies that are sensitive to cisplatin when combined with a second or third cytotoxic agent. Attempts to develop platinum compounds to reduce the side effects of cisplatin have resulted in the introduction of carboplatin and oxaliplatin. However, both drugs have proven to have inferior response rates to cisplatin especially in lung cancer. Other cytotoxic agents such as taxanes (paclitaxel, docetaxel), gemcitabine, vinorelbine, pemetrexed, and irinotecan have also been used as substitutes of cisplatin; none of these has demonstrated superior efficacy to cisplatin in lung cancer. This study represents the first time a drug has improved on cisplatin’s response rate in non-squamous NSCLC, the largest subtype of lung cancers.

Dr. George Stathopoulos and his collaborators announced exciting data from a randomized Phase III study against non-squamous non-small cell lung cancer (ns- NSCLC) mainly representing adenocarcinomas of the lung using Lipoplatin™. The results of this trial were published in October 2011 in Cancer Chemother Pharmacol. Vol 68, pages 945-950 (Open access at ). This study used Lipoplatin in combination with paclitaxel as first line treatment against ns-NSCLC and compared response rates and toxicities to a similar group of patients treated with cisplatin plus paclitaxel. This study has demonstrated an increase in tumor response rate in the Lipoplatin arm (59.22% of patients) versus the cisplatin arm (42.42%, of patients) that was statistically significant (p value = 0.036). Most major toxicities of cisplatin, especially nephrotoxicity were also reduced in the group of patients treated with Lipoplatin.

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Median survival times were 10 months for the Lipoplatin arm and 8 months for the cisplatin arm, with a p-value of 0.155. The median duration of response was 7 months for the Lipoplatin arm and 6 months for the cisplatin arm. Although not statistically significant, these results suggest the potential for superior overall survival (OS) for Lipoplatin compared to cisplatin, a hypothesis that is being tested in a larger trial. Furthermore, among the responders to Lipoplatin a subgroup of patients demonstrated a substantially higher overall survival than a comparable subgroup of cisplatin responders. After 10 months, 30% of patients in the Lipoplatin arm, as compared with just 16% of patients in the cisplatin arm, were without disease progression. By the end of the trial, there were 32 patients alive, 21 from the Lipoplatin arm (20.39%) and 11 from the cisplatin arm (11.11%). Thus, after 18 months, the number of surviving patients was approximately double for Lipoplatin versus cisplatin. Determining the gene expression profile of patients responsive to Lipoplatin is an important project; prediction of this group of patients from gene expression profiling in peripheral blood lymphocytes might result in fast track regulatory approvals by FDA and EMA.

The clinical development of Lipoplatin in adenocarcinomas establishes this drug as the most active platinum drug with significantly lower side effects.

Other Clinical Studies

Regulon has obtained the consent of EMA for a registrational Phase III study (~880 patients) with a Lipoplatin plus Alimta vs Cisplatin plus Alimta as first-line treatment of non-squamous NSCLC. This study has commenced in over 50 oncology centers across 10 EU countries, and is expected to recruit patients from USA centers of excellence as well as from oncology centers in Asian countries.

Lipoplatin monotherapy

Dr. George Stathopoulos demonstrated that Lipoplatin monotherapy against adenocarcinomas of the lung can have very high efficacy (38% partial response,

43% stable disease) with only minimal (Grade I) toxicity applied as second-line chemotherapy (after failure of the recommended first-line treatment).

It is anticipated that the success of this treatment will be even higher when applied as first line.

A total of 21 patients (2 patients 1 st -line, 10 as 2 nd -line and 9 as 3 rd -line) were treated in this study.
All 21 patients were evaluable for toxicity. Grade 1 myelotoxicity in two (9.52%) patients. Grade 1 nausea and vomiting in 4 (19.05%) patients. Grade 1 fatigue and peripheral neuropathy in 3 (14.29%) patients. No alopecia.
During the time of the drug infusion, temporary myalgia was observed in 5 patients, but it lasted for only 5-10 min.
Notably, no renal toxicity was detected, even after the 6th treatment course.
The monotherapy study shows significant response rate of Lipo/Nanoplatin in
NSCLC mostly applied as second- and third-line. A partial response of 38% with
43% SD as second-line chemotherapy is considered significant rarely seen with other drugs.

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Because this is the best result in monotherapy among 1,000 FDA-approved drugs Lipoplatin monotherapy can be applied to lung cancer as first-line treatment

Monotherapy with Lipoplatin vs other drugs




(2 and


3 -line)

PR 38%

SD 43% PD 19%

Grade 1 myelotoxicity, 9.5%

Grade 1 nausea and vomiting,


Grade 1 fatigue and peripheral neuropathy, 14%

temporary myalgia, 24% Renal toxicity, 0% Neuropathy, 0%

Stathopoulos et al. Oncol Lett.

2012 4:1013-1016. c/articles/PMC3499504/pdf/ol-


Cisplatin in

NSCLC (1 st line)

PR 11% median survival 7.6 months;

One-year survival


Grades ¾: Nausea and vomiting 21% and 19%; renal

2%; neurotoxicity 8.6%

Sandler et al, 2000 J Clin

Oncol 18:122-130.

Oxaliplatin in colorectal

PR 10% as

2 nd


PR 18% -


24% as 1



Grade 3 neuropathy in 13% Grade 3 neutropenia in 5.2% Grade 3 thrombopenia in 7.9% Grade 3/4 vomiting in 7.9% Grade 3 diarrhea in 2.6%

Becouarn and Rougier 1998

Semin Oncol. 25: 23-31.

Bécouarn et al 1998 J Clin

Oncol. 1998 16: 2739-44.

SPI-077 in NSCLC (liposomal cisplatin of SEQUUS/ ALZA/J&J)

PR 4.5%

Grade 1,2 anemia 81% Grade 1,2 nausea 38% Grade 3,4 nausea 7.7% Grade 3 itching 3.8% Grade 1,2 rash 15.3%

White et al, 2006 Br J Cancer

95, 822-828 al/v95/n7/full/6603345a.html

Avastin in ovarian cancer

PR 16%

Grade 3 to 4: hypertension (9.1%), proteinuria (15.9%), GI perforations (11.4%), arterial thromboembolic events (6.8%), deaths (6.8%), bleeding (2.3%), wound- healing complications (2.3%)

Cannistra et al, J Clin Oncol.

2007 25:5180-6

Kyprolis (Onyx) for multiple myeloma (Carfilzomi b is a modified tetrapeptid yl epoxide)

PR 18%

Grade 3 and 4 (Serious) adverse reactions: 45%. Fatique, 56%; Anemia 47%; Nausea, 45%; Thrombocytopenia, 36%; Dyspnea, 35%; Diarrhea 33%; Pneumonia, 10%; Acute renal failure, 4%; Congestive heart failure, 3%

Zangari et al 2011 Eur J Haematol. 86:484-7.

Cisplatin monotherapy induces to patients much higher toxicities (Grade 3, 4) compared to 0% Grade 3, 4 after Lipoplatin treatment. In addition, cisplatin displays a much lower efficacy than Lipoplatin in monotherapy studies (11.1%

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for cisplatin vs 38.1% for Lipoplatin). These facts are summarized in the Table that follows. Cisplatin is considered to be the best drug or the “Queen of Chemotherapy” among approximately 1,000 oncology products approved by FDA and EMA. Cisplatin has also a very broad spectrum against the vast majority of human cancers of epithelial origin (About 80-90% of cancers are epithelial malignancies). The fact that Lipoplatin displays such a big difference in efficacy compared to cisplatin advocates for the value of this drug in cancer management.

Toxicities and response

Cisplatin monotherapy

Lipoplatin monotherapy

Grades 3/4 hematologic toxicities



Grades 3/4 Neutropenia and thrombocytopenia

4.5% and 3.6%


Grades 3/4 Anemia (low hematocrit)



Grades 3/4 febrile neutropenia



Grades 3/4 Nausea and vomiting

21% and 19%


Grades 3/4 renal toxicity



Grades 3/4 Neurotoxicity



Overall response rate




Sandler AB, Nemunaitis J, Denham C, et al: Phase III trial of gemcitabine plus cisplatin versus cisplatin alone in patients with locally advanced or metastatic non-small-cell lung cancer. J Clin Oncol

18:122-130, 2000.

Stathopoulos GP, Stathopoulos J, Dimitroulis J. Two consecutive days of treatment with liposomal cisplatin in non-small cell lung cancer. Oncol Lett.

2012, vol 5:1013-1016.

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Lipoplatin Monotherapy with low-dose radiation

Clinical studies pioneered by Professor Koukourakis at the University Hospital of Alexandroupolis using fractions of radiation therapy (RT) in combination with Lipoplatin against gastric cancer patients have shown up to 80% complete response. Details of the study: Patients with locally advanced gastric cancer. Lipoplatin weekly
120 mg/m 2 (D1). 5-FU weekly 400 mg/m 2 (D1). Radiotherapy at 3.5-Gy fractions on
4 of 5 patients had complete response after 5 weekly cycles (Koukourakis et al, 2010
Int J Radiation Oncology Biol Phys 78, 150-155).
This was the first clinical demonstration of a very high efficacy of the combination of Lipoplatin + RT. Although the study included 12 patients it is hoped to stimulate oncologists to apply this treatment in larger trials.
A similar study by the same group has been extended to NSCLC using Lipoplatin monotherapy + RT with excellent efficacy and low toxicity.
A Ph.D. Thesis in Canada has shown that Lipoplatin has the best synergistic effect with radiation therapy in cell culture or in animals against glioblastomas (brain tumors) compared to other platinum drugs (cisplatin, oxaliplatin, carboplatin). The explanation is that Lipoplatin increases the intracellular uptake of the drug and the damage to the cell is much higher; then concomitant treatment with radiation enhances the damage to the point where the cancer cell is unable to repair and commits apoptotic death.
Application of Lipoplatin on Day 1 and Day 2 on cancer patients (to divide the dose and avoid side effects) and its combination with low-dose radiation therapy (1 Gy on Day 2) enhances 14 times the radiosensitizing potential of Lipoplatin as shown in preclinical studies in Canada. In these studies, Lipoplatin™ and other platinum drugs were tested on the F98 glioma cells for their ability to improve the cell uptake and increase the synergic effect when combined with ionizing radiation. Lipoplatin was shown to have the best radiosensitizing potential among all platinum compounds on F98 glioma cells. After 4 h exposure with platinum compounds, cells were irradiated (1.5 to 6.6 Gy) with a 60Co source. Lipoplatin compared to cisplatin improved the cell uptake by 3-fold because of its liposomal nature, and its radiosensitizing potential was enhanced by 14-fold ( Charest et al, 2010) .

Charest et al (2010) Concomitant treatment of F98 glioma cells with new liposomal platinum compounds and ionizing radiation. J Neurooncol. 97: 187–193.

When a similar protocol is being applied to human cancer patients the efficacy of Lipoplatin is expected to increase by a factor of 14 by combining with low-dose radiation. Thus, the new treatment protocol is providing to the medical community a drug many times more efficacious than the queen of chemotherapy, cisplatin, and without side effects. This is bringing a true revolution in cancer treatment.
Lipoplatin could become the drug of choice for all three major human cancers (lung, breast, prostate) but also in a number of smaller cancer indications. Furthermore, because of low toxicity could find application against pediatric tumors. Its liposomal nature enhances penetration through the blood-brain barrier for brain and spinal cord tumors whereas targeting of the bone marrow by liposomes makes our drug an excellent candidate for childhood and adult leukemias.

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In the image below, a 95% tumor reduction of a high-grade osteosarcoma was obtained when the patient was treated with dose-intense Lipoplatin monotherapy and fractionated radiation over a 25-week period. The patient had failed first and second- line chemotherapy. Our treatment was void of toxicities.

The images show PET/CT scans before (left) and after (right) Lipoplatin - radiation therapy in a patient with high-grade osteosarcoma. A significant lower metabolic activity of the mass can be seen consistent with approximately 95% response. Osteosarcoma, a bone cancer most

commonly seen in adolescents and young adults, is usually a high-grade malignancy treated

with four “old” drugs, namely methotrexate, doxorubicin (Adriamycin), cisplatin, and ifosfamide that cause severe side effects. Unfortunately, the past 30 years have witnessed few, if any, survival improvements. Our treatment would offer a new regiment against osteosarcomas without side effects and an amazing efficacy.

Thus, when combined with external radiation to the tumors where the nanoparticles with their toxic payload have accumulated, the result is a much better efficacy unlike any other regimens in clinical practice. With this treatment, Lipoplatin monotherapy and low-dose radiation achieve spectacular results in the management of terminal cancer patients.

Regulon’s protocol and duration of treatment

Lipoplatin is already 3-4 times more efficient than cisplatin applied as monotherapy. Combination of Lipoplatin at 200 mg/m 2 on D1,2 every 14 days or
200 mg/m 2 D1 every week for 12 weeks (3 months) with low-dose radiation therapy (2 Gy on Day 2, total dose 24 Gy in 12 weeks) enhances at least 5 times
the efficacy of Lipoplatin.
The total radiation dose administered in hospitals is 50 Gy usually given 2 Gy daily for 5 days per week and for 5 tandem weeks. The patient suffers from side effects caused by radiation. The proposed radiation dose of 24 Gy in Regulon’s protocol is without side effects because it is given over a period of 12 weeks instead of 5 weeks and is half of the recommended dose. The 24Gy over 12 weeks given in this schedule is sufficient to enhance the activity of platin already internalized by the cancer cells and enhance the killing effect of Lipoplatin in addition to the killing effect of radiation causing double strand breaks on the DNA.
This could become a universal protocol for all cancers.

Combination of Lipoplatin with energy from external source

Combination of oncothermia with Lipoplatin by the group of Dr. Shimon Slavin in Tel
Aviv has shown very high efficacy. Oncothermia (hyperthermia) increases the

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concentration of the nanoparticle in the tumor by a factor of 10-fold but also damages preferentially the tumor cells compared to the neighboring normal tissue.
The group of Dmitri Lapotko at Rice University found that gold nanoparticles, injected to animals with cancer, concentrate in tumors (just like Lipoplatin); when the animals are then irradiated with 3-6% of the standard radiation dose, gold nanoparticles absorb energy to boil the cytoplasm in their surroundings and emit secondary energy to further damage tumor cells.
Dr. Lapotko in collaboration with Regulon is now using Lipoplatin with a tremendous advantage over gold: Platinum in cisplatin is by itself the best chemotherapy drug whereas gold nanoparticles by themselves do not posses chemotherapeutic efficacy. Gold nanoparticles are also very toxic. Thus, we expect that this method will have a tremendous clinical efficacy. Laser beam radiation will cause heating of the Lipoplatin nanoparticles with release of the drug cisplatin to further damage the cancer cell.

Figure above: Schematic drawing of the methodology of Regulon and Dr. Lapotko for cancer treatment

Other developments

Lipoplatin in adrenocortical carcinoma (ACC)
Etoposide, doxorubicin, and cisplatin together with mitotane (EDP-M) have been defined as the standard treatment for late-stage disease patients with adrenocortical carcinoma. Application of Lipoplatin into anticancer cocktails tested in rats improved adrenocortical carcinoma treatment and showed the highest and sustained antitumoral effects. This study was initiated at the Ludwig-Maximilians-Universität, Munich, Germany by Drs. Hantel and Beuschlein.
Regulon has established collaboration with two eminent German clinics treating a rare cancer called adrenocortical carcinoma, ACC. A number of studies in animals were conducted and published and the first testing on patients with ACC also took place successfully in regimens incorporating Lipoplatin. The overall histological evaluation demonstrated improved off-target profiles following liposomal regimens. The first clinical data indicate improved tolerability of liposomal drugs against ACC. The German clinicians have mobilized their colleagues in MD Anderson cancer

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Center, in Italy and France and there is a global interest to apply Lipoplatin in adrenocortical carcinoma. There have been teleconferences with the consortium members and plans to apply for a European grant to support this study. Regulon has proposed Lipoplatin monotherapy with radiation to eliminate side effects, the consortium proposes LEDP-M (etoposide, liposomal doxorubicin, liposomal cisplatin, mitotane).
Lipoplatin with low-dose photodynamic therapy
Photodynamic Therapy (PDT) is a treatment for wet age-related macular degeneration (ARMD), a disease that involves abnormal blood vessel growth in the macula. The macula or macula lutea (from Latin macula, "spot" + lutea, "yellow") is an oval-shaped pigmented area near the center of the retina of the human eye. Verteporfin (trade name Visudyne), a benzoporphyrin derivative, which is a photosensitiser (i.e. light-activated) molecule, is injected intravenously into the patient. As the drug flows naturally through the body’s vascular system, it will pass into the abnormal blood vessels in the macula at the back of the eye. When stimulated by nonthermal red light with a wavelength of 689 nm in the presence of oxygen, produces highly reactive short-lived singlet oxygen and other reactive oxygen radicals, resulting in local damage to the endothelium and blockage of the vessels.
Researchers at CHUV in Lausanne have used a combination of Visudyne and Lipoplatin. Low-dose, Visudyne-mediated photodynamic therapy (photo-induction) was shown to selectively enhance tumor vessel transport causing increased uptake of systemically administered chemotherapy in various tumor types grown on rodent lungs. In a study published in 2015 they explored the efficacy of photo-induced vessel modulation combined to intravenous Lipoplatin on rodent lung tumors and the feasibility/toxicity of this approach in porcine chest cavities. Photo-induction selectively increased Lipoplatin uptake in all orthotopic tumors. It significantly increased the ratio of tumor to lung Lipoplatin concentration in sarcoma (P = 0.0008) and adenocarcinoma (P = 0.01) but not in mesothelioma, compared to IV drug application alone. Visudyne-mediated photo-induction selectively enhances the uptake of IV administered Lipoplatin in rodent lung tumors. Intrapleural VATS photo- induction with identical treatment conditions combined to IV Lipoplatin chemotherapy is feasible and well tolerated in a porcine model.
New mechanisms for killing ovarian cancer cells- Lipoplatin as an antimetastasis drug.
In a paper that appeared in September 2013 in “Gynecologic Oncology” from the “Reference Oncology Center and the National Cancer Institute” in Aviano, Italy (, the investigators, led by Dr. Donatella Aldinucci have examined the effectiveness of Lipoplatin in cisplatin- resistant cervical cancer cells. This paper was part of a Ph.D. thesis by Dr. Casagrande on new mechanisms for killing cancer cells by Lipoplatin. The submitted article won the Italian Prize as the best work of the year.
Cisplatin is the FDA recommended treatment for cervical cancer in combination with
radiation therapy. However, after initial success with cisplatin in front line chemotherapy tumors become resistant to platinum drugs.
In the Aldinucci study, Lipoplatin was effective in both cervical cancer and cisplatin- resistant cervical cancer cells thus opening the possibility to apply Lipoplatin successfully against cervical cancer both as first and second-line. Furthermore, the same study has elucidated novel mechanisms on how Lipoplatin kills cervical cancer cells:

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1. Lipoplatin treatment induced apoptosis in these cells, as evaluated by Annexin-V staining and DNA fragmentation, caspases 9 and 3 activation, Bcl-2, Bcl-xL down- regulation, and Bax up-regulation.
2. Lipoplatin inhibited the activity of Thioredoxin reductase (TrxR) in cervical cancer cells. TrxR is a selenoenzyme which is over-expressed in many tumor cells and contributes to drug resistance; this finding revealed one of the several mechanisms Lipoplatin can kill cells resistant to platinum.
3. Lipoplatin induced an increase in Reactive Oxygen Species (ROS) even in the presence of the ROS scavenger N-Acetylcysteine (NAC). This is an additional mechanism for mediating tumor cell killing by Lipoplatin.
4. Lipoplatin reduced the expression of EGFR (epidermal growth factor receptor) and its phosphorylated form. EGFR is overexpressed in many tumor cells and thus the reduction of EGFR molecules on the cell membrane by Lipoplatin would have a strong anticancer effect.
5. Lipoplatin inhibited both migration and invasion of cervical cancer cells. This is a very important feature of Lipoplatin because migration and invasion are essential steps used by cancers to mediate their metastases; over 90% of cancer patients succumb because of complications from metastases rather than the primary tumor.
Previous studies have suggested an antiangiogenesis potential of Lipoplatin from its ability to attack endothelial cells of tumor vasculature. Cisplatin, the drug inside the liposomal capsule, is the queen of classical chemotherapy with its ability to attack epithelial malignancies (more than 50% of human cancers are epithelial in origin). From the recent work of Aldinucci we conclude that the Lipoplatin nanoparticle is a candidate magic bullet for cancer: chemotherapy, antiangiogenesis and antimetastases properties all wrapped up together in the same nanoparticle.

Pharmaco-economic benefits

Lipoplatin has significant pharmaco-economic benefits: (i) it is being administered on an outpatient basis and without pre- or post-hydration compared to cisplatin treatment that requires 2 days of hospitalization. (ii), there is less use of antiemetics and of the expensive hemotopoietic factors EPO and G-CSF with Lipoplatin, compared to cisplatin, treatment. (iii) The absence of Grade III-IV nephrotoxicity in Lipoplatin also eliminates the expensive hemodialysis treatments for life required for a percentage of patients treated with cisplatin and causing permanent damage to their kidneys. (iv), because of its lower neuropathy and improvement in Quality Of Life (QOL) it remains as the drug of choice. (v) Because of the low cumulative toxicity, Lipoplatin has been administered to over 27 doses to a single patient compared to maximum 6 doses allowed for cisplatin and thus, Lipoplatin has application as a maintenance therapy to extend the life span of cancer patients.

Lipoplatin can be administered to patients with renal insufficiency

In an article published in the “Journal of Drug Delivery & Therapeutics”, Dr. George Stathopoulos and collaborators from the Errikos Dunant Hospital in Athens, Greece announce significant results from the application of Lipoplatin to patients with renal insufficiency.

In this study, 42 patients mainly with lung and bladder cancers with renal insufficiency (creatinine levels 1.6 to 4.0 mg/dl) were treated with Lipoplatin monotherapy or with its combination with gemcitabine, paclitaxel, or 5-fluorouracil-

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leucovorin. Whereas most chemotherapy regimens, especially those containing cisplatin, increase serum creatinine because of renal toxicity, Lipoplatin did not cause any increase in creatinine levels in any of the patients treated. Only Grade 1-2 myelotoxicity was observed which was mild and did not necessitate the use of growth factors. Complete response was observed in 5 patients with bladder cancer, partial response in 15 patients (8 with bladder, 2 with NSCLC and 5 with GI tract cancers) and stable disease was observed in 14 patients (3 with bladder, 6 with NSCLC and 5 with GI tract cancers). Only 8 of 42 patients had progressive disease (all 8 had NSCLC).

This work open the possibility for the use of Lipoplatin in many cancer indications where patients have enhanced creatinine and urea levels either as a results of prior chemotherapy or because of renal insufficiency from other drugs or causes.

For additional information contact:

Dr. Teni Boulikas

Chief Executive Officer

Regulon, Inc. and Regulon AE

Apollonos 1 and Varis Koropiou Avenue

Industrial Zone Koropi 19400 Attiki


Mobile: +30 6937 255 067

Skype: teni.boulikas

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