Automotive cyber-physical systems are in transition from the closed-systems to open-networking systems. As a result, in-vehicle networks such as the controller area network (CAN) have become essential to connect to inter-vehicle networks through the various rich interfaces. Newly exposed security concerns derived from this requirement may cause in-vehicle networks to pose threats to automotive security and driver’s safety. In this paper, to ensure a high level of security of the in-vehicle network for automotive CPS, we propose a novel lightweight and practical cyber defense platform, referred to as CANon (CAN with origin authentication and non-repudiation), to be enabled to detect cyber-attacks in real-time. CANon is designed based on the hierarchical approach of centralized-session management and distributed-origin authentication. In the former, a gateway node manages each initialization vector and session of origin-centric groups consisting of two more sending and receiving nodes. In the latter, the receiving nodes belonging to the given origin-centric group individually perform the symmetric key-based detection against cyber-attacks by verifying each message received from the sending node, namely origin authentication, in real-time. To improve the control security, CANon employs a one-time local key selected from a sequential hash chain (SHC) for authentication of an origin node in a distributed mode and exploits the iterative hash operations with randomness. Since the SHC can constantly generate and consume hash values regardless of their memory capacities, it is very effective for resource-limited nodes for in-vehicle networks. In addition, through implicit key synchronization within a given group, CANon addresses the challenges of a key exposure problem and a complex key distribution mechanism when performing symmetric key-based authentication. To achieve lightweight cyber-attack detection without imposing an additive load on CAN, CANon uses a keyed-message authentication code (KMAC) activated within a given group. The detection performance of CANon is evaluated under an actual node of Freescale S12XF and virtual nodes operating on the well-known CANoe tool. It is seen that the detection rate of CANon against brute-force and replay attacks reaches 100% when the length of KMAC is over 16 bits. It demonstrates that CANon ensures high security and is sufficient to operate in real-time even on low-performance ECUs. Moreover, CANon based on several software modules operates without an additive hardware security module at an upper layer of the CAN protocol and can be directly ported to CAN-FD (CAN with Flexible Data rate) so that it achieves the practical cyber defense platform.