Enhancing Anti-tumor Efficacy by Combination of WK0202 with Immune Checkpoint Inhibitors and Immunogenic Cell Death Inducer including Chemotherapy/Targeted Therapy

( Sei Hoon Yang ),( Hong-seob So ),( Sun Rock Moon ),( Tae Hwan Kwak ),( Hong Seob So ) 대한결핵 및 호흡기학회 2021 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.129 No.0

Background/Aim Immune checkpoint inhibitors (ICIs) including anti-PD1, anti-PD-L1, and anti-CTLA-4 show promising therapeutic outcomes. However, ICI has a limited effect in some carcinomas. To overcome these shortcomings, ICI was combined with chemotherapy, targeted therapy or radiation therapy inducing immunogenic cell death (ICD). ICD involves the release of damage-associated molecular patterns (DAMPs) from dying tumor cells to activate tumor-specific immune responses. ICD also causes the release of tumor-associated antigens, destruction of physical barriers, and promoting immune cell re-penetration, which trigger adaptive immune cells to elicit an antitumor immune response. However, nonetheless, if the intratumoral environment surrounding cancer cells with high concentrations of lactate, TGF-b, HIF1/2, and VEGF, which favors cancer growth and metastasis, does not change, these therapeutic outcomes will be limited. WK0202, our drug candidate, does not directly induce ICD, but fundamentally changes the tumor microenvironment favorable for cancer growth and metastasis by modulating aerobic glycolysis. Therefore, our research aim is to demonstrate that the combination of WK0202 with ICD stimulation such as chemotherapy or targeted therapy and ICIs could induce even more anti-tumor efficacy. Methods WK0202, which is currently completed Phase 1 study by NADIANBIO Ltd., will be administered in combination with an immunotherapy in Phase 2 clinical trial. Using a preclinical mouse lung cancer model system, we examined the anti-tumor efficacy of triple combination consisted of ICI, ICD inducer, and WK0202. Anti-PD1 was used as ICI, and paclitaxel/carboplatin was used as ICD inducer. Results WK0202, when administered in combination with ICD and ICI, produced significantly higher anticancer effects than when each component was administered alone or when ICI and ICD were combined. Conclusions Therefore, our research suggests a possibility that the combination of WK0202 with ICD stimulation such as chemotherapy or targeted therapy and ICIs could induce even more clinical benefits in various cancer patients.

CT26 고형암을 내포하는 BALB/cKorl Syngeneic 마우스에서 Ecklonia cava의 항암효과 및 항염증효과

노유정,김지은,진유정,설아윤,송희진,김태렬,민경선,박은서,박기호,황대연 한국생명과학회 2023 생명과학회지 Vol.33 No.11

염증반응(inflammation)은 발병, 진행, 악성 전이를 포함한 암의 진행과정(tumorigenesis)에서 중요한 역할을 수행하기 때문에 암 치료를 위한 전략으로 고려되고 있다. 감태(Ecklonia cava) 열수추출물(AEC)의 항암활성 동안 나타나는 항염증 반응을 연구하기 위하여, 비만세포(mast cells)의 분포, inducible nitric oxide synthase (iNOS)단백질, cyclooxygenase-2 (COX-2)단백질, nuclear factor (NF)-κB단백질, inflammasome 구성 단백질, inflammatory cytokines 발현의 변화는 AEC를 5주간 경구투여한 CT26 대장암을 내포하는 BALB/ cKorl syngeneic 마우스에서 분석하였다. AEC를 처리한 후, 고형암의 무게와 조직 절편의 괴사 부위가 vehicle처리그룹에 비하여 감소하였다. 비만세포의 수는 vehicle처리그룹에 비하여 AEC처리그룹에서 증가했지만 COX-2와 iNOS의 발현은 AEC처리그룹에서 감소하였다. 또한, NF-κB, NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC)과 Caspase-1 (Cas-1)단백질의 발현도 유사한 감소가 관찰되었다. 더불어, tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α)와 interleukin-6 (IL-6)의 mRNA 발현이 vehicle처리그룹에 비하여 AEC처리그룹에서 감소하였다. 이러한 결과는 AEC가 CT26 고형암을 내포하는 BALB/cKorl syngeneic 마우스에서 항암활성은 염증반응과 밀접한 관련이 있음을 제시하고 있다. The inflammatory response have been considered as one of important targets for cancer treatment because they play a key role during all steps of tumor development including initiation, promotion, malignant conversion and progression. To investigate the anti-inflammatory response during anti-tumor activity of an aqueous extracts of Ecklonia cava (AEC), alterations on the distribution of mast cells and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor (NF)-κB, inflammasome compositional protein and inflammatory cytokines were examined in CT26 colon tumor-bearing BALB/cKorl syngeneic mice after administrating AEC for five weeks. After treatment of AEC, total weight of tumor and necrotic region of tumor section were significantly decreased compared to vehicle treated group. The number of infiltered mast cells was higher in AEC treated group than vehicle treated group, while the expression levels of COX-2 and iNOS were decreased in AEC treated group. Also, similar decrease pattern were detected in the expression levels of NF-κB, NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase-1 (Cas-1) after AEC treatment although the decrease rate was varied. Furthermore, the mRNA expressions of three inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α) and interleukin-6 (IL-6) were remarkably decreased in AEC treated group compared to vehicle treated group. These results suggest that inhibition of inflammatory response may be tightly associated with anti-tumor activity of AEC in CT26 colon tumor-bearing BALB/cKorl syngeneic mice.

Anti-tumor Effects of Penfluridol through Dysregulation of Cholesterol Homeostasis

Wu, Lu,Liu, Yan-Yang,Li, Zhi-Xi,Zhao, Qian,Wang, Xia,Yu, Yang,Wang, Yu-Yi,Wang, Yi-Qin,Luo, Feng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

Background: Psychiatric patients appear to be at lower risk of cancer. Some antipsychotic drugs might have inhibitory effects on tumor growth, including penfluridol, a strong agent. To test this, we conducted a study to determine whether penfluridol exerts cytotoxic effects on tumor cells and, if so, to explore its anti-tumor mechanisms. Methods: Growth inhibition of mouse cancer cell lines by penfluridol was determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxic activity was determined by clonogenic cell survival and trypan blue assays. Animal tumor models of these cancer cells were established and to evaluate penfluridol for its anti-tumor efficacy in vivo. Unesterified cholesterol in cancer cells was examined by filipin staining. Serum total cholesterol and tumor total cholesterol were detected using the cholesterol oxidase/p-aminophenazone (CHOD-PAP) method. Results: Penfluridol inhibited the proliferation of B16 melanoma (B16/F10), LL/2 lung carcinoma (LL/2), CT26 colon carcinoma (CT26) and 4T1 breast cancer (4T1) cells in vitro. In vivo penfluridol was particularly effective at inhibiting LL/2 lung tumor growth, and obviously prolonged the survival time of mice bearing LL/2 lung tumors implanted subcutaneously. Accumulated unesterified cholesterol was found in all of the cancer cells treated with penfluridol, and this effect was most evident in LL/2, 4T1 and CT26 cells. No significant difference in serum cholesterol levels was found between the normal saline-treated mice and the penfluridol-treated mice. However, a dose-dependent decrease of total cholesterol in tumor tissues was observed in penfluridol-treated mice, which was most evident in B16/F10-, LL/2-, and 4T1-tumor-bearing mice. Conclusion: Our results suggested that penfluridol is not only cytotoxic to cancer cells in vitro but can also inhibit tumor growth in vivo. Dysregulation of cholesterol homeostasis by penfluridol may be involved in its anti-tumor mechanisms.

Effects of canine mesenchymal stem cells-derived exosomes in a mouse model of canine mammary tumor

정소연(So Yeon Jeong),이세아(Se-A Lee),구나연(Na-Yeon Gu),이지현(Jienny Lee),이윤희(Yoon-Hee Lee),현방훈(Bang-Hun Hyun) 한국예방수의학회 2020 예방수의학회지 Vol.44 No.4

Canine mammary tumors account for ~30% of all tumors in the female dogs and approximately 50% of the tumors are malignant. Exosomes have been the focus of great interest, as they appear to be involved in numerous important cellular processes. In this study, we examined the anti-tumor effects of canine mesenchymal stem cells-derived exosomes (MSC-exosomes) in an experimental murine mammary tumor model using canine mammary carcinoma cells, REM134. The MSC-exosomes were injected tumor site and tail vein of REM134 xenografted mice. We found that tumor size of the MSC-exosomes-treated group decreased compared to those of the only tumor group in REM134-driven tumorigenic mouse model. In addition, the MSC-exosomes-treated tumor group showed meaningfully reduced expression levels of the MMP-3, IL-1β, IL-6, and TNF-α compared to those in the tumor group. Specifically, we confirmed that the expression level of the CD133, potent cancer stem cell (CSC) markers, decreased in the MSC-exosomes-treated tumor group compared to the tumor group. This study suggests that the MSC-exosomes exhibited anti-tumor effects through downregulating CSC-related markers in the canine mammary tumor murine model. Further study is needed in the future, and we are conducting research on the detailed anti-tumor mechanism of the MSC-exosomes.

사매가 위암세포에 미치는 영향

류봉하,김진성,윤상협,류기원,홍상선,Ryu Bong-Ha,Kim Jin-Sung,Yoon Sang-Hyub,Ryu Ki-Won,Hong Sang-Sun 대한한방내과학회 2003 大韓韓方內科學會誌 Vol.24 No.2

Background : Nowadays many researches about it s cure are going on world widely since cancer is one of the most human health threatening diseases. In Chinese and North Korean medicine, Duchesnea india(Audra.) Foche. is practically used to treat many kinds of cancer, but in Korea it is rarely used. So, we need to scientifically identify anti-tumor effects of Duchesnea india(Audra.) Foche. Objective : We are aimed to identify anti-tumor effects of Duchesnea india(Audra.) Foche. on the stomach cancer cells through molecular biologic methods. Material & Methods : We used AGS as stomach cancer cells from American Type Culture Collection. We added the boiled extract of Duchesnea india(Audra.) Foche. $5{\mu}l$(Sample I), $10{\mu}l$(Sample II) to cultural media(ml)for 0,6, 12, 24, 48 hours. We measured the killing effect on stomach cancer cells through Tryphan blue exclusion test and the suppressive effect on viability of stomach cancer cells via MTT assay. the quantitative RT-PCR was used to examine their effect on the revelation of Bcl-2, Bcl-XL, and Bax, which are genes related to apoptosis. We measured change of mitochondria membrane permeability and membrane potential via flow cytometry. Result : 1. The killing effect on stomach cancer cells showed that each test groups killed more stomach cancer cells than the control group with a time(6 hours later) and density dependent manner, which was statistical significance. 2. The suppressive effect on viability of stomach cancer cells showed that each test groups had more suppressive effects on viability of stomach cancer cells than the control group with a time(6 hours later), which was statistical significance. 3. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 and Bcl-XL decreased with a density manner which was statistical significance. but the revelation of Bax was not changed with statistical significance. 4. As a result of this test, Duchesnea india(Audra.) Foche. caused apoptosis by decreasing the absorbance of mitochondria with statistical significance. and also induced apoptosis by decreasing the membrane potential of mitochondria. Conclusion : This experiment showed that Duchesnea india(Audra.) Foche. has anti-tumor effect with statistical significance. This is in vitro experiment and basic experiment on Duchesnea india(Audra.) Foche. We hope more progressive researchs on Duchesnea india(Audra.) Foche. will go on and its anti-tumor effects will be more practically identified.

강황이 수종의 암세포에 미치는 영향

윤주호,김진성,윤상협,류봉하,Yoon, Joo-Ho,Kim, Jin-Sung,Yoon, Sang-Hyub,Ryu, Bong-Ha 대한한방내과학회 2006 大韓韓方內科學會誌 Vol.27 No.2

Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

순기화중탕(順氣和中湯)과 Doxorubicin의 병용이 Colon-26의 항암효과에 미치는 영향

신민규,김봉석,오중한,임희용,김동우,최빈혜,변준석,Shin, Min-Kyu,Kim, Bong-Suk,Oh, Jung-Han,Lim, Hee-Yong,Kim, Dong-Woo,Choi, Bin-Hye,Byun, Joon-Seok 대한한방내과학회 2004 大韓韓方內科學會誌 Vol.25 No.2

In order to evaluate the anti-tumor and synergic effect of Soonkiwhajungtang with doxorubicin, the inhibitory concentration(IC), IC50 and IC90 of single use of doxorubicin and Soonkiwhajungtang with their concomitant treatment against Colon-26(Murine Rectum Carcinoma) was observed using MTT(Microculture Tetrazolium test) assay. In addition, their anti-tumor effects were also observed in the xenograft nude mice models against 3LL cell lines. Soonkiwhajungtang may only mimic direct anti-tumor effects against 3LL cell lines, but signs of worsening induced by implantation of tumor cell lines generally decreased, while the total WBC and lymphocyte numbers increased. Therefore, experimentation suggests that Soonkiwhajungtang extracts reduced the critical toxicity of doxorubicin, and that Soonkiwhajungtang extracts have favorable synergic effects when combined with doxorubicin.

순기화중탕(順氣和中湯)과 Doxorubicin의 병용이 Colon-26의 항암효과에 미치는 영향

신민규,김봉석,오중한,임희용,김동우,최빈혜,김상찬,변준석,Shin Min-Kyu,Kim Bong-Suk,Oh Jung-Han,Lim Hee-Yong,Kim Dong-Woo,Choi Bin-Hye,Kim Sang-Chan,Byun Joon-Seok 대한한의학방제학회 2004 大韓韓醫學方劑學會誌 Vol.12 No.2

In order to evaluate the anti-tumor and synergic effect of Soonkiwhajungtang with doxorubicin, the inhibitory concentration(IC), $IC_{50}\;and\;IC_{90}$ of single use of doxorubicin and Soonkiwhajungtang with their concomitant treatment against Colon-26(Murine Rectum Carcinoma) was observed using MTT(Microculture Tetrazolium test) assay. In addition, their anti-tumor effects were also observed in the xenograft nude mice models agianst to Colon-26 cell lines. Soonkiwhajungtang has only mimic direct anti-tumor effect against to Colon-26 cell lines but they were decreased general depressed signs induced by implantation of tumor cell lines and increased the total WBC and lymphocyte numbers. So, it is considered or expected that Soonkiwhajungtang extracts were reduced the critical toxicity of doxorubicin and shows favorable synergic effect with doxorubidn and Soonkiwhajungtang extracts.

보중익기합대칠기탕(補中益氣合大七氣湯)과 Doxorubicin의 병용이 3LL의 항암효과에 미치는 영향

이윤희,김봉석,오중한,임희용,김동우,최빈혜,김상찬,변준석,Lee Yun-Hee,Kim Bong-Suk,Oh Jung-Han,Lim Hee-Yong,Kim Dong-Woo,Choi Bin-Hye,Kim Sang-Chan,Byun Joon-Seok 대한한의학방제학회 2004 大韓韓醫學方劑學會誌 Vol.12 No.1

In order to evaluate the anti-tumor and synergic effect of Bojungikkeehapdaechilkitang with doxorubicin, the inhibitory concentration(IC), $IC_{50}\;and\;IC_{90}$ of single use of doxorubicin and Bojungikkeehapdaechilkitang with their concomitant treatment against 3LL(Lewis lung carcinoma) was observed using MTT(Microculture Tetrazolium test) assay. In addition, their anti-tumor effects were also observed in the xenograft nude mice models agianst to 3LL cell lines. Bojungikkeehapdaechilkitang has only mimic direct anti-tumor effect against to 3LL cell lines but they were decreased general depressed signs induced by implantation of tumor cell lines and increased the total WBC and lymphocyte numbers. So, it is considered or expected that Bojungikkeehapdaechilkitang extracts were reduced the critical toxicity of doxorubicin and shows favorable synergic effect with doxorubicin and Bojungikkeehapdaechilkitang extracts.

보중익기합대칠기탕(補中益氣合大七氣湯)과 Doxorubicin의 병용이 MKN-45의 항암효과에 미치는 영향

이윤희,김봉석,오중한,임희용,김동우,최빈혜,변준석,Lee Yun-Hee,Kim Bong-Suk,Oh Jung-Han,Lim Hee-Yong,Kim Dong-Woo,Choi Bin-Hye,Byun Joon-Seok 대한한방내과학회 2004 大韓韓方內科學會誌 Vol.25 No.1

In order to evaluate the anti-tumor and synergic effect of BoJungIkKeeHapDaeChilKi-Tang on doxorubicin, the inhibitory concentration(IC), IC50 and IC90 of single use of doxorubicin and BoJungIkKeeHapDaeChilKi-Tang with their concomitant treatment against MKN-45(Human stomach carcinoma) was observed using MTT(Microculture Tetrazolium test) assay. In addition, their anti-tumor effects were also observed in the xenograft nude mice models agianst MKN-45 cell lines. BoJungIkKeeHapDaeChilKi-Tang has only mimic direct anti-tumor effect against to MKN-45 cell lines but they were decreased general depressed signs induced by implantation of tumor cell lines and increased the total WBC and lymphocyte numbers. So, it is considered or expected that BoJungIkKeeHapDaeChilKi-Tang extracts were reduced by the critical toxicity of doxorubicin and shows favorable synergic effect with doxorubicin and BoJungIkKeeHapDaeChilKi-Tang extracts.