瘤内注射E1B缺失腺病毒(H101)与化疗联合治疗恶性肿瘤的Ⅱ期临床试验

背景及目的:具有瘤内复制能力的病毒(或称溶瘤病毒)用于治疗恶性肿瘤的研究近年来发展相当迅速,其中E1B缺失腺病毒是目前研究进展最快的溶瘤病毒之一。本研究采用自身对照的方法,观察在全身化疗的同时,部分瘤内注射E1B缺失腺病毒(H101)的近期疗效和不良反应。方法:采用多中心开放式研究。共入选50例,在全身化疗的同时,对目标病灶给予H101瘤内注射,每天1次,每次1支(0.5ml,含5×1011病毒颗粒),连续5天为1疗程。治疗期间同时联合常规化疗。记录疗效并与未注射H101的自身瘤灶对比,同时观察不良反应。结果:在可评价疗效的46例患者中,注射病灶的总有效率为30.4%,而按ITT人群分析有效率为28.0%(14/50),其中CR为3例,PR为11例;对照病灶的总有效率为13.0%,其中CR为1例,PR为5例。注射病灶的有效率明显高于对照病灶(P<0.001)。主要不良反应为注射部位局部疼痛(26.9%)和发热(30.2%),有4例出现Ⅰ级肝功能异常,1例出现Ⅱ级肝功能异常,1例出现Ⅳ级肝功能异常,4例出现Ⅳ级血液学毒性。结论:在全身化疗的同时,瘤内注射E1B缺失腺病毒(H101)对多种不同类型的恶性肿瘤具有一定疗效,其毒副作用较低,易为患者接受。     

瘤内注射E1B缺失腺病毒治疗恶性肿瘤的Ⅱ期临床研究

目的 :观察瘤内注射E1B缺失腺病毒注射液 (H10 1)治疗恶性肿瘤的近期疗效和不良反应。方法 :采用多中心开放式研究。符合入选标准共 53例 ,给予H 10 1瘤内注射 ,每天一次 ,每次一支 (5× 10 11病毒颗粒数 /0 5ml) ,连续 5天为一疗程。治疗期间及治疗后观察并记录疗效和不良反应。结果 :在可评价疗效的 49例患者中 ,CR 2例 ,PR 9例 ,MR 8例 ,SD 19例 ,PD 11例 ,总有效率为 2 2 .4% (11/49) ,按ITT人群分析有效率为 2 0 .8%(11/53 ) ,临床获益患者 (CR PR MR SD)占 77.6% (3 8/49)。主要不良反应为注射局部疼痛 (57 4% )和发热(3 5 2 % ) ,未见严重不良事件。结论 :E1B缺失腺病毒 (H10 1)瘤内注射对不同类型的恶性肿瘤具有一定疗效 ,其毒副作用较低 ,易为病人接受     

溶瘤腺病毒的研究进展

 引言受某些肿瘤患者在感染病毒后出现自发性肿瘤缓解这一现象的启发,早在上个世纪初便有溶瘤病毒治疗的研究[1] 。腺病毒发现后不久也曾被用来治疗头颈部恶性肿瘤,注射腺病毒后肿瘤有不同程度缩小,但治疗后肿瘤易复发,效果难以持久;直到1996年,Bischoff等[2 ] 首次报道去除部分E1B的重组腺病毒Onyx 0 15能在p5 3异常的肿瘤细胞选择性复制引起肿瘤杀伤作用,溶瘤腺病毒研究才再次受到广泛关注并且发展迅速,出现了许多新型溶瘤腺病毒种类,使溶瘤腺病毒成为恶性肿瘤一种新的治疗平台。1　溶瘤腺病毒的作用机理[1,3- 5]许多种DNA病毒,如腺病毒、SV4 0、HPV 16等都可以改变宿主细胞细胞周期,其原因主要是病毒产生一些蛋白作用于宿主细胞细胞周期调节蛋白,使静止期细胞进入细胞周期以利于病毒DNA的复制。不同病毒影响细胞周期的机制不一,腺病毒主要通过Rb和p5 3细胞信号通路干扰宿主细胞细胞周期。1.1　Rb和p5 3细胞信号通路(见图1) :Rb(眼癌蛋白,retinoblastomaprotein)的作用是与E2F(使细胞从G1期进入S期的一种转录因子)结合,细胞内游离E2F水平下降,细胞由...     

E1B缺失腺病毒H101的研究进展

 E1B缺失腺病毒H101是采用基因重组技术对人5型腺病毒进行基因重组得到的一种溶瘤腺病毒，它主要是删除了人5型腺病毒的E1B-55×103和E3区19×103基因片段，具有在肿瘤细胞中特异性复制而最终导致溶瘤的特性。     

基因工程腺病毒(H101)瘤内注射联合化疗治疗头颈部及食管鳞癌的Ⅲ期临床研究

背景与目的：H101是利用基因重组技术得到的一种删除了E1B部分基因片段的溶瘤腺病毒,其在一定剂量范围内对肿瘤有明显的抑制作用。本研究目的是比较H101瘤内注射联合顺铂(c platin,DDP) 5-氟尿嘧啶(5-fluorouracil,5-FU)或阿霉素(adriamycin,ADM) 5-FU方案化疗与单纯化疗对头颈、食管鳞癌的治疗效果及毒副作用。方法：160例入选患者,根据过去化疗史分别选定不同化疗方案。未用过PF方案化疗或用过PF方案有效者采用PF方案化疗,已用过PF方案无效者则采用AF方案化疗。采用多中心随机对照分组,分别加用(A组)或不加用(B组)肿瘤浅表病灶内注射H101。两组均为21天一个周期,所有患者至少接受2个周期的治疗。结果：完全符合方案标准的病例123例,A1组(PF H101)有效率为78．8％,B1组(PF only)为39．6％,A2组(AF H101)为50％(7／14),B2组(AF only)50％(2／4)。A1、B1组间以及A1 A2、B1 B2组间疗效差异有显著性(P=0．000)。45．7％病例出现轻～中度发热,28．3％出现注射局部反应,9．8％出现流感样症状。结论：H101注射液瘤内局部注射联合化疗的客观有效率比单纯化疗组高,显示H101瘤内注射对于头颈、食管鳞癌具有明确的治疗作用,且具有较高的安全性。     

柯萨奇病毒腺病毒受体的表达对基因工程腺病毒抗瘤活性的影响

背景与目的:腺病毒是研究最多的一种溶瘤病毒,也是基因治疗最常用的一种载体,但作为溶瘤病毒或载体,其治疗恶性肿瘤的疗效很大程度上受到组织器官表达柯萨奇病毒腺病毒受体(Coxsackieandadenovirusreceptor,CAR)的影响。本研究旨在观察CAR表达对腺病毒感染力及临床疗效的影响。方法:收集29例基因工程腺病毒(H101)临床试验患者的病理标本,用免疫组织化学法检测CAR表达,分析CAR表达与临床疗效的关系。体外通过流式细胞仪检测不同肿瘤细胞表面CAR表达,用MTT法测定病毒对不同细胞的抑制率,以观察CAR表达与H101感染力的关系。结果:29例恶性肿瘤患者中,10例有效病例CAR阳性表达者7例(70.0%),19例无效病例CAR阳性表达仅6例(31.6%),CAR表达与临床疗效呈显著性相关(P=0.048)。体外实验结果显示,不同肿瘤细胞株CAR的表达存在差异,其表达量与H101对细胞的感染力(病毒对肿瘤细胞抑制率)呈正相关(r=0.986)。结论:CAR的表达与腺病毒的感染力及临床疗效密切相关,CAR高表达的肿瘤对腺病毒治疗的疗效较低表达者明显为好。     

H101溶瘤腺病毒联合长春瑞滨/顺铂一线治疗晚期非小细胞肺癌

目的:探讨溶瘤腺病毒H101瘤内注射联合长春瑞滨/顺铂(NP)治疗晚期非小细胞肺癌(NSCLC)的疗效与安全性。方法:病理学或细胞学确诊的初治NSCLC患者随机接受经皮肺穿剌瘤内注射H1011.5×1012病毒颗粒联合NP方案化疗(A组)或单纯NP方案化疗(B组)。治疗2个周期后进行一次评价疗效,随访无进展生存时间(TTP)和生存期。结果:A组19例可评价患者中,总体疗效部分缓解(PR)5例,疾病稳定(SD)10例,疾病进展(PD)4例;B组可评价17例中,PR3例,SD9例和PD5例。第1次评价疗效时,A组PD1例,而B组PD5例,B组治疗失败率显著高于A组(P<0.05)。2组生存曲线几乎相似,但A组6月、9月和1年生存率均要稍高于B组;A组中位TTP时间也较B组有所延长。A组副反应中,除了非感染发热外,其他不良反应与B组相似。A组有2例发生轻度气胸。结论:经皮肺穿剌瘤内注射H101联合NP方案治疗晚期NSCLC是可行、安全与有效的。     

溶瘤病毒H101结合局部加热治疗对转移肿瘤病灶的远端效应

背景与目的:远端效应是肿瘤的局部治疗引起的远处抗肿瘤作用。本研究探索溶瘤腺病毒H101局部注射结合局部加热治疗诱发远端抗瘤效应的可行性。方法:放化疗失败或拒绝放化疗并有转移病灶的5例肿瘤患者(2例鼻咽癌,1例肺癌,1例骨旁质肉瘤和1例膀胱癌),以H101(5×1011～15×1011VP)瘤内注射联合60min42℃局部热疗进行治疗。结果:2例患者注射灶、非注射灶肿瘤完全消退而长期生存,3例患者也观察到非注射病灶有不同程度缓解后死亡,生存期分别为29、15、13个月。结论:H101瘤内注射联合加热治疗能诱发一定的远端抗瘤效应。     

基因工程腺病毒(H101)瘤内注射联合化疗治疗鼻咽癌的疗效观察

目的:观察瘤内注射基因工程腺病毒(H101)联合化疗治疗鼻咽癌的近期疗效和不良反应方法:符合入选标准的鼻咽癌患者54例,给予H101瘤内注射,每天一次,每次一支(5×1011病毒颗粒/0.5ml),连续5天,同时联合PF方案(DDP 20mg/m2静滴,d1～5,5-FU 500mg/m2静滴,d1～5)或AF方案(ADM 50mg/m2静注d1;5-FU 500mg/m2静滴,d1～5)化疗,21天为一周期,所有患者均接受2个周期的治疗.结果:在可评价的54例患者中注射病灶的总有效率为77.7%,其中CR 10例,PR 32例.主要不良反应为注射部位局部疼痛20.4%,非感染性发热40.7%,流感样症状9.3%,未见严重不良事件.结论:在全身化疗的同时瘤内注射基因工程腺病毒(H101)对鼻咽癌有明确的治疗作用,不良反应低,易为患者接受.     

调控食管癌癌胚抗原基因表达对溶瘤腺病毒H101抗癌作用的影响

目的 研究调控食管癌癌胚抗原(CEA)基因表达对溶瘤腺病毒H101抗癌作用的影响,探讨影响H101敏感性的内在因素.方法 选择CEA低表达的EC9706细胞株(EC9706-SCEA)和CEA过表达的EC9706细胞株(EC9706-CEA)复苏培养,细胞生长实验检测调控EC9706细胞CEA基因表达对细胞生长的影响,通过细胞毒性实验和裸鼠体内实验检测H101对不同CEA表达水平EC9706细胞的杀伤效果.结果 细胞生长实验表明,EC9706-SCEA、EC9706-CEA和EC9706细胞群体倍增时间分别为(30.9±2.0)h、(31.1±2.5)h和(29.1±2.6)h,差异无统计学意义(P＞0.05).细胞毒性实验显示,当感染复数(MOI) ≥0.01 PFU时,H101对EC9706-SCEA细胞的抑制率明显高于EC9706、EC9706-CEA、EC9706-siCon和EC9706 -Con细胞(P＜0.05).不同时间各组裸鼠移植瘤体积测定显示,H101对治疗组裸鼠皮下移植瘤的生长均有抑制作用,但其对EC9706-SCEA治疗组移植瘤生长的抑制作用(抑瘤率为61.5％ ～ 74.5％)显著强于EC9706治疗组(抑瘤率为35.5％ ～44.8％)和EC9706-CEA治疗组(抑瘤率为32.3％～38.5％),差异有统计学意义(p＜0.05).结论 溶瘤病毒H101对EC9706-SCEA细胞的杀伤力明显增强.沉默CEA基因的表达,有望成为提高溶瘤病毒H101抗癌效果的新途径.     

重组人内皮抑素腺病毒注射液Ⅰ期临床耐受性试验

背景与目的:血管生成抑制剂在抗肿瘤新生血管生成方面显示出良好的临床应用前景,已有多个Ⅰ期临床试验研究人内皮抑素重组蛋白的安全性和抗瘤活性.本试验目的是确定携带人内皮抑素基因的重组腺病毒注射液(Ad-Es)的最大耐受剂量,并推荐Ⅱ期临床试验的用量和用法.方法:设预试验组1例,1×1010病毒颗粒,单次瘤内注射.普通试验组遵循以一个自然对数级的1/2的方式递增,共14例,分三个剂量组:1×1011、5×1011、1×1012病毒颗粒/次,瘤内注射,每周1次,连续2周.结果:未观察到剂量限制性毒性,各受试者均显示出良好的耐受性.主要的不良事件为:局部反应和发热.其他不良反应有轻微的肝功能异常和流感样症状,如头痛、肌痛、乏力等.1例鼻咽癌放疗后鼻咽复发并颏下淋巴结转移的患者病情好转,12例病情稳定,2例出现进展.结论:人体对Ad-Es耐受性良好,晚期肿瘤患者使用1×1012病毒颗粒/次,瘤内注射,每周1次,连续2周,初步观察到Ad-Es的抗瘤活性.推荐Ⅱ期临床给药剂量为1.0×1012病毒颗粒/次,每周1次,连续4周.     

Phase II trial of intratumoral administration of ONYX-015, a replication-selective adenovirus, in patients with refractory head and neck cancer.

PURPOSE: To determine the safety, humoral immune response replication, and activity of multiple intratumoral injections of ONYX-015 (replication selective adenovirus) in patients with recurrent squamous cell carcinoma of the head and neck (SCCHN). PATIENTS AND METHODS: This phase II trial enrolled patients with SCCHN who had recurrence/relapse after prior conventional treatment. Patients received ONYX-015 at a dose of 2 x 10(11) particles via intratumoral injection for either 5 consecutive days (standard) or twice daily for 2 consecutive weeks (hyperfractionated) during a 21-day cycle. Patients were monitored for tumor response, toxicity, and antibody formation. RESULTS: Forty patients (30 standard and 10 hyperfractionated) received 533 injections of ONYX-015. Standard treatment resulted in 14% partial to complete regression, 41% stable disease, and 45% progressive disease rates. Hyperfractionated treatment resulted in 10% complete response, 62% stable disease, and 29% progressive disease rates. Treatment-related toxicity included mild to moderate fever (67% overall) and injection site pain (47% on the standard regimen, 80% on the hyperfractionated regimen). Detectable circulating ONYX-015 genome suggestive of intratumoral replication was identified in 41% of tested patients on days 5 and 6 of cycle 1; 9% of patients had evidence of viral replication 10 days after injection during cycle 1, and no patients had evidence of replication > or = 22 days after injection. CONCLUSION: ONYX-015 can be safely administered via intratumoral injection to patients with recurrent/refractory SCCHN. ONYX-015 viremia is transient. Evidence of modest antitumoral activity is suggested.     

Selective replication and oncolysis in p53 mutant tumors with ONYX-015, an E1B-55kD gene-deleted adenovirus, in patients with advanced head and neck cancer: a phase II trial

ONYX-015 is an E1B-55kDa gene-deleted adenovirus engineered to selectively replicate in and lyse p53-deficient cancer cells. To evaluate the selectivity of ONYX-015 replication and cytopathic effects for the first time in humans, we carried out a Phase II clinical testing of intratumoral and peritumoral ONYX-015 injection in 37 patients with recurrent head and neck carcinoma. Patients received ONYX-015 at a daily dose of 1 x 10(10) plaque-forming units (pfu) via intratumoral injection for 5 days during week 1 of each 3-week cycle (n = 30; cohort A), or 1 x 10(10) pfu twice a day for 10 days during weeks 1 and 2 of each 3-week cycle. Posttreatment biopsies documented selective ONYX-015 presence and/or replication in the tumor tissue of 7 of 11 patients biopsied on days 5-14, but not in immediately adjacent normal tissue (0 of 11 patients; P = 0.01). Tissue destruction was also highly selective; significant tumor regression (>50%) occurred in 21% of evaluable patients, whereas no toxicity to injected normal peritumoral tissues was demonstrated. p53 mutant tumors were significantly more likely to undergo ONYX-015-induced necrosis (7 of 12) than were p53 wild-type tumors (0 of 7; P = 0.017). High neutralizing antibody titers did not prevent infection and/or replication within tumors. ONYX-015 is the first genetically engineered replication-competent virus to demonstrate selective intratumoral replication and necrosis in patients. This agent demonstrates the promise of replication-selective viruses as a novel therapeutic platform against cancer.     

Transrectal gene therapy of the prostate in the canine model

Direct transrectal delivery of therapeutic genes utilizing adenoviral vectors for advanced prostate cancer may offer effective treatment at the molecular level. Large animal models to assess feasibility and the intraprostatic and systemic dissemination patterns of these vectors have not been reported. For these studies, a replication-deficient (E1(-)/E3(-)) recombinant adenovirus (AdRSVlacZ) expressing bacterial beta-galactosidase (beta-gal) was delivered under transrectal ultrasound guidance. Two prostate biopsies, followed by concurrent injection of 4.8 x 10(9) pfu of the adenoviral vector divided into either 1 or 2 mL of diluent, were performed (n=4). Swabs of the rectum, sputum, and urine were collected and after 72 hours, the animals were sacrificed. Specimens were assayed for the presence of virus and beta-gal activity. Rectal swabs were transiently positive, whereas urine and sputum samples showed no detectable vector throughout the experiment. Beta-gal activity was observed at the prostate injection sites with detectable activity noted up to 7.5 mm away from the injection site. Systemic dissemination was observed regardless of the injected volume. In conclusion, transrectal prostate biopsy with concurrent prostate injection is a feasible method to deliver therapeutic adenoviral vectors for the treatment of prostate cancer; however, systemic distribution and temporary rectal shedding of virus should be anticipated.     

A phase I study of Onyx-015, an E1B attenuated adenovirus, administered intratumorally to patients with recurrent head and neck cancer.

An E1B 55 kDa gene-deleted adenovirus, Onyx-015, which reportedly selectively replicates in and lyses p53-deficient cells, was administered by a single intratumoral injection to a total of 22 patients with recurrent head and neck cancer. The objectives of this Phase I study were to determine the safety, feasibility, and efficacy of this therapy and determine any correlation to p53 status. Six cohorts were investigated with a dose escalation from 10(7)-10(11) plaque-forming units. Toxicity was assessed using NCIC criteria. Tumor response was assessed by clinical and radiological measurement. Blood samples were taken to detect adenovirus DNA and neutralizing antibody to adenovirus. Tumor biopsies were taken to detect adenovirus by in situ hybridization. Treatment was well tolerated, with the main toxicity being grade 1/2 flu-like symptoms. Dose-limiting toxicity was not reached at the highest dose of 10(11) plaque-forming units. Twenty-one of the 22 patients treated showed an increase in neutralizing antibody to adenovirus. In situ hybridization showed viral replication in 4 of 22 patients treated, all of whom had mutant p53 tumors. Using conventional response criteria, no objective responses were observed. However, magnetic resonance imaging scans were suggestive of tumor necrosis at the site of viral injection in five patients, three of whom were classified using nonconventional criteria as partial responders, and two of whom were classified using nonconventional criteria as minor responders. Of these five cases, four had mutant p53 tumors. The response duration for the three partial responders was 4, 8, and 12 weeks. An additional eight patients had stable disease in the injected tumors lasting from 4-8 weeks. These preliminary results show that intratumoral administration of Onyx-015 is feasible, well tolerated, and associated with biological activity. Further investigation of Onyx-015, particularly with a more frequent injection protocol and in combination with systemic chemotherapy, is warranted.